Gamma-aminobutyric Acid Shunt Research Articles

A multi-omics study revealed the effect of pulsed light treatment on germinated brown rice: Promotion of sprouting efficiency and gamma-aminobutyric acid enrichment

Physical fields are increasingly proposed in the production of germinated grain foods, thus shorting the preparation cycle of high value-added sprouted grain foods. Here, pulsed light treatment (PLT) with 300 times at a single energy of 400 J was found to significantly promote the shoot length and gamma-aminobutyric acid (GABA) content in 8 varieties of germinated brown rice (GBR). Functional annotation analysis showed that the brown rice germination was promoted by the biosynthesis of phenylalanine, carbohydrate metabolism, and energy metabolism pathway. Moreover, PLT influenced the genes and metabolites involved in the GABA shunt and the polyamine degradation pathway, promoting the accumulation of GABA. Transcription factor OsbZIP56 was closely related to the transcript level of regulatory gene for germination and GABA enrichment. Furthermore, overexpression of OsbZIP56 in rice significantly promoted germination and GABA accumulation. These findings would be significant for improving the germination efficiency of edible seeds, promoting accumulation of active ingredients, and exploring gene functions in food materials.

The reprogramming of mitochondrial energy metabolism in Rice: The role of the TCA cycle and GABA shunt under Arsenic exposure

AbstractArsenic (As) pollution is hazardous for all living organisms. Plants growing in As‐contaminated soil are less performant. Recently, seed priming has attracted significant attention for its beneficial effects on plants. Numerous studies have associated priming technique with the induction of stress memory against traumatic conditions, but its relation to crop's energy balance is under‐researched. The present study highlights variations in energy metabolic pathways, particularly TCA (tricarboxylic acid) cycle and GABA (gamma‐aminobutyric acid)‐shunt in multi‐nutrient [silicon (Si), iron (Fe), zinc (Zn), and ascorbic acid (AA)] primed rice seedlings under As stress. Arsenic exposure decreased the TCA cycle enzymes' activity while intensifying GABA accumulation and metabolism. This enhancement in GABA shunt provides an alternative energy source for As‐stressed seedlings. However, priming and further Fe‐supplementation reduced As‐toxicity by accelerating the activity of both energy pathways, namely, TCA and GABA cycle. Improved energy regulation by priming and Fe‐augmentation also enhanced the functionality of anabolic respiratory‐interactive‐pathway, i.e., carbon‐nitrogen metabolism. The As‐tolerance response of priming and Fe‐augmentation was further analyzed from the expression profiling of nitrogen and phosphorous utilization genes. Out of 10 genes, particular enhancement was observed in OsNiR, OsGS, and OsGOGAT expression with improved macro‐micronutrient content of the seedlings. Thus, irrespective of As infestations, multi‐nutrient (Si, Fe, Zn, and AA) seed priming approach along with Fe‐supplementation may emerge as a beneficial strategy to improve plant metabolic capacity in terms of nitrogen, phosphorous, and carbon assimilation to support normal vigor because they balance cellular energy levels.

Metabolic Role of GABA in the Secretory Function of Pancreatic β-Cells: Its Hypothetical Implication in β-Cell Degradation in Type 2 Diabetes.

The stimulus-secretion coupling of a glucose-induced release is generally attributed to the metabolism of the hexose in the β-cells in the glycolytic pathway and the citric acid cycle. Glucose metabolism generates an increased cytosolic concentration of ATP and of the ATP/ADP ratio that closes the ATP-dependent K+-channel at the plasma membrane. The resultant depolarization of the β-cells opens voltage-dependent Ca2+-channels at the plasma membrane that triggers the exocytosis of insulin secretory granules. The secretory response is biphasic with a first and transient peak followed by a sustained phase. The first phase is reproduced by a depolarization of the β-cells with high extracellular KCl maintaining the KATP-channels open with diazoxide (triggering phase); the sustained phase (amplifying phase) depends on the participation of metabolic signals that remain to be determined. Our group has been investigating for several years the participation of the β-cell GABA metabolism in the stimulation of insulin secretion by three different secretagogues (glucose, a mixture of L-leucine plus L-glutamine, and some branched chain alpha-ketoacids, BCKAs). They stimulate a biphasic secretion of insulin accompanied by a strong suppression of the intracellular islet content of gamma-aminobutyric acid (GABA). As the islet GABA release simultaneously decreased, it was concluded that this resulted from an increased GABA shunt metabolism. The entrance of GABA into the shunt is catalyzed by GABA transaminase (GABAT) that transfers an amino group between GABA and alpha-ketoglutarate, resulting in succinic acid semialdehyde (SSA) and L-glutamate. SSA is oxidized to succinic acid that is further oxidized in the citric acid cycle. Inhibitors of GABAT (gamma-vinyl GABA, gabaculine) or glutamic acid decarboxylating activity (GAD), allylglycine, partially suppress the secretory response as well as GABA metabolism and islet ATP content and the ATP/ADP ratio. It is concluded that the GABA shunt metabolism contributes together with the own metabolism of metabolic secretagogues to increase islet mitochondrial oxidative phosphorylation. These experimental findings emphasize that the GABA shunt metabolism is a previously unrecognized anaplerotic mitochondrial pathway feeding the citric acid cycle with a β-cell endogenous substrate. It is therefore a postulated alternative to the proposed mitochondrial cataplerotic pathway(s) responsible for the amplification phase of insulin secretion. It is concluded the new postulated alternative suggests a possible new mechanism of β-cell degradation in type 2 (perhaps also in type 1) diabetes.

Arbuscular mycorrhiza differentially adjusts central carbon metabolism in two contrasting genotypes of Vigna radiata (L.) Wilczek in response to salt stress

The study aimed at investigating Arbuscular Mycorrhiza (AM) mediated metabolic changes in two genotypes of mungbean (Vigna radiata) differing in their salt tolerance in presence of salt stress (100 mM NaCl). Colonisation by Claroideoglomus etunicatum resulted in higher growth, photosynthetic efficiency, total protein content, and lower levels of stress markers, indicating alleviation of stress in mungbean plants. AM differentially upregulated the components of Tricarboxylic acid (TCA) cycle in salt tolerant (ST) and salt sensitive (SS) genotypes that could be correlated to AM-mediated moderation in nutrient uptake. Under salt stress, while maximum increase in the activity of α-ketoglutarate dehydrogenase (65%) was observed in mycorrhizal (M)-ST; the increase in isocitrate dehydrogenase (79%) and fumarase (133%) activities was maximum in M-SS plants over their non-mycorrhizal (NM) counterparts. Apart from TCA, AM also affected gamma-aminobutyric acid (GABA) and glyoxylate pathways. Activities of enzymes implicated in GABA shunt increased in both the genotypes under stress resulting in increase in GABA concentration (46%). Notably, glyoxylate pathway was induced by AM in SS only, wherein M-SS exhibited significantly higher isocitrate lyase (49%) and malate synthase (104%) activities, reflected in higher malic acid concentration (84%), than NM under stress. The results suggest that AM moderates the central carbon metabolism and strategizes towards boosting the formation of stress-alleviating metabolites such as GABA and malic acid, especially in SS, bypassing the steps catalysed by salt-sensitive enzymes in TCA cycle. The study, therefore, advances the understanding on mechanisms by which AM ameliorates salt stress.

Enhancing the resistance of peach fruit against Monilinia fructicola using exogenous nitric oxide by activating the gamma-aminobutyric acid shunt

Nitric oxide (NO) can induce plant disease resistance through signal transduction. The gamma-aminobutyric acid (GABA) shunt possesses different response modes for resisting different pathogens. However, the effect of the GABA shunt in NO-induced plant disease resistance is not clear. Herein, the role of the GABA shunt in the resistance of peach fruit induced by NO against Monilinia fructicola infection was investigated together with possible interaction pathways. NO treatment repressed the disease of peach fruit, increased the endogenous NO content, and enhanced the activities and gene expression of GABA shunt enzymes within 48 h, including glutamate decarboxylase (GAD), GABA transaminase (GABA-T) and succinic semialdehyde dehydrogenase (SSADH). However, the NO scavenger 2-(4-carboxyphenyl)− 4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) aggravated the disease of peach fruit, and inhibited the activities and gene expression of GABA shunt enzymes. The above results highlighted the importance of the GABA shunt in NO-induced disease resistance of peach fruit. It was noteworthy that the nitrosylated level of GAD4, GABA-T3 and SSADH was up-regulated at 6 and 12 h in NO-treated fruit compared with the control, which might be related to the enhanced GABA shunt activity. Additionally, NO treatment up-regulated the expression of glutamate receptor (PpGLR) and glutamate dehydrogenase (PpGDH), promoted Ca2+ influx, strengthened the activity of the tricarboxylic acid cycle (TCAC), and raised the ATP content and energy charge. Overall, the data indicated that NO treatment induced peach fruit disease resistance by activating the GABA shunt.

The role of GABA in islet function.

Gamma aminobutyric acid (GABA) is a non-proteinogenic amino acid and neurotransmitter that is produced in the islet at levels as high as in the brain. GABA is synthesized by the enzyme glutamic acid decarboxylase (GAD), of which the 65 kDa isoform (GAD65) is a major autoantigen in type 1 diabetes. Originally described to be released via synaptic-like microvesicles or from insulin secretory vesicles, beta cells are now understood to release substantial quantities of GABA directly from the cytosol via volume-regulated anion channels (VRAC). Once released, GABA influences the activity of multiple islet cell types through ionotropic GABAA receptors and metabotropic GABAB receptors. GABA also interfaces with cellular metabolism and ATP production via the GABA shunt pathway. Beta cells become depleted of GABA in type 1 diabetes (in remaining beta cells) and type 2 diabetes, suggesting that loss or reduction of islet GABA correlates with diabetes pathogenesis and may contribute to dysfunction of alpha, beta, and delta cells in diabetic individuals. While the function of GABA in the nervous system is well-understood, the description of the islet GABA system is clouded by differing reports describing multiple secretion pathways and effector functions. This review will discuss and attempt to unify the major experimental results from over 40 years of literature characterizing the role of GABA in the islet.

The synergistic effects of gamma-aminobutyric acid and salinity during the enhancement of microalgal lipid production in photobioreactors

In the present work, combined application of gamma-aminobutyric acid (GABA) and salinity was used to stimulate biolipid production in the microalga Ankistrodesmus sp. EHY in a vertical tubular photobioreactor. Under 2.5 g L−1 NaCl combined with 50 μM GABA, the strain showed 1.36- and 1.27-fold increases in lipid content (59.42%) and lipid productivity (235.13 mg L−1 d−1), respectively, compared with the control. Wide-ranging analyses of the energy yield, biodiesel quality, cellular ultrastructure, signalling molecules, and lipogenesis genes were conducted to elucidate the role of GABA plus salinity on algal lipid accumulation. By using GABA and salinity treatments, the triacylglycerol (TAG) content and energy yield were increased, whereas the carbohydrate yield was decreased. The biodiesel quality of the algal biomass-derived lipids met the approved standards. The combinational strategy also increased the levels of endogenous reactive oxidative species (ROS), Ca2+, an autophagy-related protein (ATG8), glutathione (GSH) and proline, which might be associated with the increase in lipid synthesis and salt stress tolerance. Moreover, transcriptomic and metabolomic data indicated that the expression levels of TAG assembly, antioxidative system and autophagy-related genes, and the metabolite levels of the TCA cycle, GABA shunt and several stress hormones were consistent with the enhancements in lipid accumulation and stress tolerance. In the presence of GABA and salinity, exogenous proline (50 μM) and salicylic acid (SA; 12.5 mg L−1) further increased the lipid productivity to 280.48 mg L−1 d−1 and 263.58 mg L−1 d−1, respectively. Overall, the results demonstrate the significance, mechanism and applicability of using exogenous GABA combined with salinity to effectively stimulate bioenergy production by microalgae.

Exogenous methyl jasmonate enhances phytochemicals and delays senescence in harvested strawberries by modulating GABA shunt pathway

In order to illustrate the relationship between methyl jasmonate (MeJA) and gamma aminobutyric acid (GABA) in maintaining the quality and improving the postharvest life of strawberry fruit, the harvested fruit were treated with MeJA for 16 h at 20 °C and stored at 3 ± 0.5 °C for 12 days. MeJA enhanced the expression levels of GABA shunt pathway-related genes, including glutamate decarboxylase, GABA transaminase, and succinic semialdehyde dehydrogenase, leading to an increase in GABA accumulation. Treated fruit showed higher levels of total acids, anthocyanins, total phenolics, antioxidants, and phenylalanine ammonia-lyase activity, and lower levels of cell membrane deterioration, total soluble solids, polyphenol oxidase activity and decay incidence rate. The results suggest that the positive effects of MeJA in extending the fruit postharvest life, enhancing phytochemical compounds, and decreasing the decay incidence rate may be due to the effects on motivating GABA shunt pathway and PAL enzyme activity.

Characterization of GABA-Transaminase Gene from Mulberry (Morus multicaulis) and Its Role in Salt Stress Tolerance.

Gamma-aminobutyric acid (GABA) has been reported to accumulate in plants when subjected to salt stress, and GABA-transaminase (GABA-T) is the main GABA-degrading enzyme in the GABA shunt pathway. So far, the salt tolerance mechanism of the GABA-T gene behind the GABA metabolism remains unclear. In this study, the cDNA (designated MuGABA-T) of GABA-T gene was cloned from mulberry, and our data showed that MuGABA-T protein shares some conserved characteristics with its homologs from several plant species. MuGABA-T gene was constitutively expressed at different levels in mulberry tissues, and was induced substantially by NaCl, ABA and SA. In addition, our results demonstrated that exogenous application of GABA significantly reduced the salt damage index and increased plant resistance to NaCl stress. We further performed a functional analysis of MuGABA-T gene and demonstrated that the content of GABA was reduced in the transgenic MuGABA-T Arabidopsis plants, which accumulated more ROS and exhibited more sensitivity to salt stress than wild-type plants. However, exogenous application of GABA significantly increased the activities of antioxidant enzymes and alleviated the active oxygen-related injury of the transgenic plants under NaCl stress. Moreover, the MuGABA-T gene was overexpressed in the mulberry hairy roots, and similar results were obtained for sensitivity to salt stress in the transgenic mulberry plants. Our results suggest that the MuGABA-T gene plays a pivotal role in GABA catabolism and is responsible for a decrease in salt tolerance, and it may be involved in the ROS pathway in the response to salt stress. Taken together, the information provided here is helpful for further analysis of the function of GABA-T genes, and may promote mulberry resistance breeding in the future.

EP013: Novel biomarkers of succinic semialdehyde dehydrogenase deficiency highlight opportunities for screening and detection of GABA catabolism pathway abnormalities

GABA (gamma-aminobutyric acid) is catabolized in the mitochondrial matrix via the GABA shunt, a 2-step enzymatic pathway which encompasses the transamination of GABA to succinic semialdehyde by GABA-transaminase (GABA-T), followed by oxidation to succinate by succinic semialdehyde dehydrogenase (SSADH). Deficiency of SSADH results in accumulation of GABA and reduction of succinic semialdehyde to gamma-hydroxybutyrate (GHB), the principal neurotoxin in SSADH deficiency (SSADHD). SSADHD is an autosomal recessive disorder associated with biallelic pathogenic variants in ALDH5A1, typically characterized by developmental delay, hypotonia, intellectual disability, ataxia, seizures, hyperkinetic behavior, aggression, psychiatric disorders, and sleep disturbances.

Blockage of citrate export prevents TCA cycle fragmentation via Irg1 inactivation.

The metabolism of activated macrophages relies on aerobic glycolysis, while mitochondrial oxidation is disrupted. In lipopolysaccharide-activated macrophages, the citrate carrier (CIC) exports citrate from mitochondria to enhance glycolytic genes through histone acetylation. CIC inhibition or Slc25a1 knockdown reduces the occupancy of H3K9ac to hypoxia-inducible factor-1α (HIF-1α) binding sites in promoters of glycolytic genes to restrain glycolysis. HIF-1α also transcriptionally upregulates immune-responsive gene 1 for itaconate production, which is inhibited by CIC blocking. Isotopic tracing of [U-13C6] glucose shows that CIC blockage prevents citrate accumulation and itaconate production by reducing glycolytic flux and facilitating metabolic flux in the TCA cycle. Isotopic tracing of [U-13C5] glutamine reveals that CIC inhibition reduces succinate accumulation from glutaminolysis and the gamma-aminobutyric acid shunt by enhancing mitochondrial oxidation. By restraining glycolysis, CIC inhibition increases NAD+ content to ensure mitochondrial biogenesis for oxidative phosphorylation. Furthermore, blockage of citrate export reduces cerebral thrombosis by inactivation of peripheral macrophages.

Metabolic responses of sugar beet to the combined effect of root hypoxia and NaCl-salinity

The combined occurrence of salt stress and hypoxia leads to increased growth reduction and severe toxic effects compared to salt stress alone. In the present work, we analyzed the metabolic response of sugar beet (Beta vulgaris L.) to salt stress combined with hypoxia in roots as well as in young and mature leaves. B. vulgaris plants were grown in a hydroponic culture under low and high salt concentrations combined with normoxic and hypoxic conditions. A non-targeted metabolic approach was used to identify the biochemical pathways underlying the metabolic and physiological adaptation mechanisms.Young and mature leaves showed a similar metabolic response to salt stress alone and combined stresses, accumulating sugar compounds. Osmoprotectants such as proline and pinitol were accumulated under combined stress. Roots exposed to hypoxic conditions showed increased TCA (tricarboxylic acid cycle) intermediates levels such as succinate, fumarate and malate. During hypoxia, the concentration of free amino acids as well as intermediates of the GABA (gamma-aminobutyric acid) shunt increased in roots as well as in leaves. The combination of salt stress and hypoxia results in a severe stress response in roots and leaves. A partial flux of the TCA cycle linked with the GABA shunt might be activated during hypoxia to regain reduction equivalents.

GABA shunt: a key-player in mitigation of ROS during stress

Unfavorable environmental conditions such as heat, cold, drought, metal/metalloid toxicity, and pathogens enhance production of intra-and inter-cellular levels of reactive oxygen species (ROS) in plants. ROS, acting as signaling molecules, activate signal transduction pathways in response to various stresses. Alternatively, ROS cause irreversible cellular damage due to lipid peroxidation, oxidation of protein, inactivation of enzymes, DNA damage, and interact with other vital constituents of plant cells through their strong oxidative properties, which drastically alter plant morphological structures, becoming disadvantageous for survival and productivity. Higher plants have complex defense systems to scavenge ROS. Being a central molecule of the defense system, gamma-aminobutyric acid (GABA) is ubiquitous from prokaryotes to eukaryotes cells. GABA helps mitigate ROS in plants and GABA shunt pathway plays a key role either as metabolites or endogenous signaling molecules in several regulatory mechanisms under stress conditions. The GABA transporters (GATs) being activated with the attachment of GABA under environmental stress stimuli facilitate high content of Ca2+ into the cytosol. Ca2+ combines with calmodulin (CaM) -binding domain that activates the glutamate decarboxylase (GAD) enzyme for the conversion of glutamate into GABA. This synchronized process regulates GABA shunt gene expressions under stress conditions and improves defense mechanisms in plants. This review highlights the regulatory aspects of GABA shunt pathway for ROS production as well as in the defense mechanism of plants.

The influence of laser beam and high light intensity on lentil (Lens culinaris) and wheat (Triticum aestivum) seedlings growth and metabolism

Light is considered as an environmental signal that controls plants growth and development. Sufficient duration, quality and wavelength are mainly responsible for switching on/off many physiological processes. The aim of this study was to examine the role of gamma aminobutyric acid (GABA) shunt pathway specific response in wheat (Triticum aestivum) and lentil (Lens culinaris) seedlings to blue LED laser (intensity= 1.48 W/cm2) and high intensity invisible ultraviolet light (intensity= 3.99 W/cm2) treatments separately, with respect to: seed germination, seedling growth, oxidative damage in term of reactive oxygen substances accumulation, GABA accumulation level, total proteins and total carbohydrates level and glutamate decarboxylase gene (GAD) expression. Data showed a remarkable changes in proteins content, carbohydrates content, chlorophyll level, dry and fresh weight, GABA accumulation, MDA level, and GAD m-RNA level under blue LED laser and high intensity invisible ultraviolet light treatments; separately in the both wheat and lentil cultivars. There were no significant differences of root emergence in all cultivars under all treatments. The concentrations of MDA increased in all cultivars under blue LED laser and high intensity invisible ultraviolet light treatments with increasing exposure durations. GABA levels increased and correlated significantly in all cultivars as the exposure duration of blue LED laser and high light intensity increased. Significant decrease in chlorophyll a and b contents in Um Qayes, Jordan1 and Jordan2 cultivars under all laser and high intensity invisible ultraviolet light treatments. A significant difference was found in proteins content in all cultivars in response to laser and high intensity invisible ultraviolet light treatments. Carbohydrates content in all cultivars used in this study were significantly decreased under laser treatments. The transcription level of GAD increased significantly under blue LED laser and high intensity invisible ultraviolet light in all cultivars used in this study. GAD expression was high in Hurani75 cultivar in agreement with the low level of GABA accumulation under high intensity invisible ultraviolet light at 60 min which strongly suggested that GABA was catabolized to enter another pathways (C:N balance) that was confirmed by increase in higher level of carbohydrates and proteins beside reduction in MDA content at the same exposure duration. On other hand, GAD expression in Jordan 2 was lower in response to laser treatment under the same exposure duration. The high increase of GAD gene expression explains the high accumulation of GABA level under both treatments. Results indicated that GABA shunt is a key metabolic pathway that allows wheat and lentil to adapt blue LED laser and high intensity invisible ultraviolet light stressful treatments.

The involvement of gamma-aminobutyric acid shunt in the endoplasmic reticulum stress response of Arabidopsis thaliana

The endoplasmic reticulum (ER) is the main site of secretory protein production and folding and its homeostasis under environmental stress is vital for the maintenance of the protein secretory pathway. The loss of homeostasis and accumulation of unfolded proteins in the ER is referred to as ER stress. Although, γ-aminobutyric acid (GABA) is an important regulator of stress response in plants, its roles during ER stress remains unclear. This study investigated the involvement of GABA in the ER stress response of plants. For this, changes in GABA metabolism under ER stress was analysed in Arabidopsis thaliana, then to study the response of the ER-folding machinery, plants were treated with exogenous GABA under ER stress. The antibiotic tunicamycin, which inhibits N-glycosylation was used to specifically induce ER stress. This stress up-regulated the expression of five glutamate decarboxylase (GAD) genes except GAD2 and GABA content of A. thaliana plants increased with an increasing concentration of tunicamycin (0.1 μg ml−1 and 0.25 μg ml−1). Moreover, expressions of genes involved in the conversion of GABA to succinate was also induced, while genes involved in transport across plasma and mitochondrial membrane showed no response to ER stress. The exogenous treatment of plants with 1-and 5-mM GABA increased plant performance under ER stress but 0.1 mM proved ineffective. Plants treated with GABA under ER stress had decreased expression of ER stress marker genes such as BIP1, BIP3 or CNX, but the expression of genes related to ER stress perception or ER-associated protein degradation showed no changes with respect to GABA treatments.

Changes in photosynthesis and respiratory metabolism of maize seedlings growing under low temperature stress may be regulated by arbuscular mycorrhizal fungi

Arbuscular mycorrhizal fungi as an important soil microbe have been demonstrated to mitigate the harmful effects of stress on plants. However, little is known about the molecular mechanisms underlying the AM symbiotic response to low temperature. Here, differentially expressed genes (DEGs) in the maize seedlings were identified after inoculating AMF under low temperature conditions. A total of 10,400 DEGs were obtained among four treatments, including non-inoculated AMF under ambient temperature (NMA), inoculated AMF under ambient temperature (MA), non-inoculated with low temperature stress (NML), and inoculated with low temperature stress (ML). The relative expression of 858 genes increased and that of 497 genes decreased in AM plants under low temperature stress. 24 DEGs were identified related to photosynthesis and respiratory metabolism. Among these DEGs, 10 genes were upregulated, and 14 genes were downregulated. The results show that inoculating AMF might decrease the production and transmission of electrons under low temperature, and the cyclic electron flow process in chloroplasts was stimulated to protect plants against low temperature. The fungi also influenced transmission of electrons and production of phosphoric acid in mitochondria in response to low temperature. CO2 assimilation capacity was affected and the tricarboxylic acid cycle was promoted by the adjustments in the glycolysis, pentose phosphate pathway, gamma-aminobutyric acid shunt pathway, and glyoxylic acid cycle to produce more adenosine triphosphate and raw materials for other metabolic pathways under low temperature. These findings provide new insight into low temperature tolerance induced by AMF, and help identify genes for further investigation and functional analyses.

The GABA shunt pathway in germinating seeds of wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) under salt stress

AbstractSoil salinity is one of the major abiotic stresses affecting seed germination, crop growth and productivity. In this study, seeds of three wheat (Triticum aestiveum L.) and three barley (Hordeum vulgare L.) cultivars were treated with different concentrations of NaCl to investigate the effect of salt on seed germination physiology and metabolism through the characterization of seed germination pattern, gamma-aminobutyric acid (GABA) shunt metabolite accumulation [GABA, glutamate (Glu) and alanine (Ala)] and glutamate decarboxylase (GAD) expression using RT-PCR. A trend of decreasing germination percentage with increasing NaCl concentrations was observed. Under all salt stress treatments, data showed significant increase with positive correlation (r = 0.50–0.99) between abundance of GABA shunt metabolites and salt concentration in all wheat and barley cultivars for 5 days. Increased GABA content was associated with a small but significant increase in Ala and Glu content in all cultivars. In all NaCl treatments, the transcription of GAD in terms of RNA abundance showed a significant increase in all cultivars with positive correlation (r = 0.50–0.98). Data showed significant association between GAD RNA transcription and the response of germinating seeds to salt stress in terms of GABA shunt metabolite accumulation. The elevated expression of GAD under salinity suggests the need for elevated activity of the GAD-mediated conversion of Glu to GABA during seed germination, which provides alternative metabolic routes to the respiratory machinery, balancing carbon and nitrogen metabolism and osmolyte synthesis in germinating seeds of wheat and barley under salt stress.

Metabolomic Dynamics Reveals Oxidative Stress in Spongy Tissue Disorder During Ripening of Mangifera indica L. Fruit.

Spongy tissue disorder, a mesocarp specific malady, severely affects the flavor and pulp characters of Alphonso mango fruit reducing its consumer acceptability. Here, we investigated comparative metabolomic changes that occur during ripening in healthy and spongy tissue-affected fruits using high resolution mass spectrometric analysis. During the spongy tissue formation, 46 metabolites were identified to be differentially accumulated. These putative metabolites belong to various primary and secondary metabolic pathways potentially involved in maintaining the quality of the fruit. Analysis revealed metabolic variations in tricarboxylic acid cycle and gamma amino butyric acid shunt generating reactive oxygen species, which causes stressed conditions inside the mesocarp. Further, reduced levels of antioxidants and enzymes dissipating reactive oxygen species in mesocarp deteriorate the fruit physiology. This oxidative stress all along affects the level of amino acids, sugars and enzymes responsible for flavor generation in the fruit. Our results provide metabolic insights into spongy tissue development in ripening Alphonso mango fruit.

Role of GABA transaminase in the regulation of development and senescence in Arabidopsis thaliana

GABA (gamma amino butyric acid) transaminase (GABA-T) shows significant regulatory function during plant developmental process. It is involved in the reduction of GABA to succinic semialdehyde and participates in GABA shunt pathway, which critically contributes to nitrogen metabolism during senescence stage of plant’s life cycle. Thus, to study the function of GABA-T gene in the development and senescence processes, Arabidopsis thaliana GABA-T knock out mutants (pop2-1 and pop2-3) were characterized by the assays of leaf senescence parameters (survival efficacy, total chlorophyll content, electrolyte leakage of membrane, and lipid peroxidation) and GABA shunt components in different developmental stages. GABA-T gene mutants (pop2-1 and pop2-3) impact the overall plant morphology, leaf development, flowering time and display early onset of senescence. It was observed that pop2 mutants declined the leaf survival efficacy, total chlorophyll, GABA, GABA-T as well as glutamate decarboxylase (GAD) enzymes activities. Knockout mutation in GABA-T gene adversely elevated the electrolyte leakage of membrane, lipid peroxidation at S3 stage (30 days old plant) rather than S5 stage (50 days old plant). ROS generation and cell death was also increased in pop2 mutant leaf earlier. Taken together, our results suggest a prominent role of GABA-T gene in leaf senescence and development processes in Arabidopsis thaliana.

Comparison of pathway analysis and constraint-based methods for cell factory design

BackgroundComputational strain optimisation methods (CSOMs) have been successfully used to exploit genome-scale metabolic models, yielding strategies useful for allowing compound overproduction in metabolic cell factories. Minimal cut sets are particularly interesting since their definition allows searching for intervention strategies that impose strong growth-coupling phenotypes, and are not subject to optimality bias when compared with simulation-based CSOMs. However, since both types of methods have different underlying principles, they also imply different ways to formulate metabolic engineering problems, posing an obstacle when comparing their outputs.ResultsIn this work, we perform an in-depth analysis of potential strategies that can be obtained with both methods, providing a critical comparison of performance, robustness, predicted phenotypes as well as strategy structure and size. To this end, we devised a pipeline including enumeration of strategies from evolutionary algorithms (EA) and minimal cut sets (MCS), filtering and flux analysis of predicted mutants to optimize the production of succinic acid in Saccharomyces cerevisiae. We additionally attempt to generalize problem formulations for MCS enumeration within the context of growth-coupled product synthesis. Strategies from evolutionary algorithms show the best compromise between acceptable growth rates and compound overproduction. However, constrained MCSs lead to a larger variety of phenotypes with several degrees of growth-coupling with production flux. The latter have proven useful in revealing the importance, in silico, of the gamma-aminobutyric acid shunt and manipulation of cofactor pools in growth-coupled designs for succinate production, mechanisms which have also been touted as potentially useful for metabolic engineering.ConclusionsThe two main groups of CSOMs are valuable for finding growth-coupled mutants. Despite the limitations in maximum growth rates and large strategy sizes, MCSs help uncover novel mechanisms for compound overproduction and thus, analyzing outputs from both methods provides a richer overview on strategies that can be potentially carried over in vivo.