Endogenous GABA Content Research Articles

SlGAD2 is the target of SlTHM27, positively regulates cold tolerance by mediating anthocyanin biosynthesis in tomato.

Cold stress significantly limits the yield and quality of tomato. Deciphering the key genes related to cold tolerance is important for selecting and breeding superior cold-tolerant varieties. γ-aminobutyric acid (GABA) responds to various types of stress by rapidly accumulating in plant. In this study, glutamic acid decarboxylase (GAD2) was a positive regulator to enhance cold stress tolerance of tomato. Overexpression of SlGAD2 decreased the extent of cytoplasmic membrane damage and increased the endogenous GABA content, antioxidant enzyme activities, and reactive oxygen species (ROS) scavenging capacity in response to cold stress, whereas Slgad2 mutant plants showed the opposite trend. In addition, SlGAD2 induced anthocyanin biosynthesis in response to cold stress by increasing the content of endogenous GABA. Further study revealed that SlGAD2 expression was negatively regulated by the transcription factor SlTHM27. However, the transcript levels of SlTHM27 were repressed under cold stress. Antioxidant enzyme activities, SlGAD2 transcript levels, GABA and anthocyanin contents were significantly increased in Slthm27 mutant plants. Further, our study demonstrated that SlTHM27 decreases SlGAD2-promoted cold resistance in tomato by repressing SlGAD2 transcription. Overall, our results showed that the SlTHM27-SlGAD2 model regulates the cold tolerance in tomato by regulating GABA and anthocyanin.

Exogenous γ-aminobutyric acid (GABA) improves salt-inhibited nitrogen metabolism and the anaplerotic reaction of the tricarboxylic acid cycle by regulating GABA-shunt metabolism in maize seedlings

Salinity stress hampers the growth of most crop plants and reduces yield considerably. In addition to its role in metabolism, γ-aminobutyric acid (GABA) plays a special role in the regulation of salinity stress tolerance in plants, though the underlying physiological mechanism remains poorly understood. In order to study the physiological mechanism of GABA pathway regulated carbon and nitrogen metabolism and tis relationship with salt resistance of maize seedlings, we supplemented seedlings with exogenous GABA under salt stress. In this study, we showed that supplementation with 0.5 mmol·L−1 (0.052 mg·g−1) GABA alleviated salt toxicity in maize seedling leaves, ameliorated salt-induced oxidative stress, and increased antioxidant enzyme activity. Applying exogenous GABA maintained chloroplast structure and relieved chlorophyll degradation, thus improving the photosynthetic performance of the leaves. Due to the improvement in photosynthesis, sugar accumulation also increased. Endogenous GABA content and GABA transaminase (GABA-T) and succinate semialdehyde dehydrogenase (SSADH) activity were increased, while glutamate decarboxylase (GAD) activity was decreased, via the exogenous application of GABA under salt stress. Meanwhile, nitrogen metabolism and the tricarboxylic acid (TCA) cycle were activated by the supply of GABA. In general, through the regulation of GABA-shunt metabolism, GABA activated enzymes related to nitrogen metabolism and replenished the key substrates of the TCA cycle, thereby improving the balance of carbon and nitrogen metabolism of maize and improving salt tolerance.

The combined application of Arabic gum coating and γ-aminobutyric acid mitigates chilling injury and maintains eating quality of ‘Kinnow’ mandarin fruits

Low temperature storage of citrus generally extends the storage potential but leads to chilling injury appearance on the rind of fruits. The said physiological disorder has been found associated with changes in the metabolism of cell walls and other attributes. In this work, the influence of Arabic gum [AG (10 %)] and γ-amminobutyric acid [GABA (10 mmol L−1)] either alone or in combined form was studied on fruits of ‘Kinnow’ mandarin during storage at 5 ± 1 °C for 60 days. The results exhibited that the combined AG + GABA treatment significantly suppressed weight loss (5.13 %), chilling injury (CI) symptoms (2.41 score), incidence of disease (13.33 %), respiration rate [(4.81 μmol kg−1 h−1) RPR] and ethylene production [(0.86 nmol kg−1 h−1) EPR]. In addition, AG + GABA application reduced relative electrolyte (37.89 %) leakage (REL), malondialdehyde [(25.99 nmol kg−1) MDA], superoxide anion [(15.23 nmol min−1 kg−1) O2•−] and hydrogen peroxide [(27.08 nmol kg−1) H2O2] along with lower lipoxygenase [(23.81 U mg−1 protein) LOX] and phospholipase D [(14.07 U mg−1 protein) PLD] enzyme activities compared with control. The AG + GABA treated ‘Kinnow’ group showed higher glutamate decarboxylase [(43.18 U mg−1 protein) GAD] and lower GABA transaminase [(15.93 U mg−1 protein) GABA-T] activity having higher endogenous GABA (42.02 mg kg−1) content. The fruits treated with AG + GABA exhibited higher cell walls (CW) components such as Na2CO3-soluble pectin [(6.55 g kg−1) NCSP], chelate-soluble pectin [(7.13 g kg−1) CSP] and protopectin [(11.03 g kg−1) PRP] concentrations along with lower water-soluble pectin [(10.64 g kg−1) WSP] compared to control. In addition, ‘Kinnow’ fruits treated with AG + GABA showed higher firmness (8.63 N) and lower activities of CW degrading such as cellulase [(11.23 U mg−1 protein) CX], polygalacturonase [(22.59 U mg−1 protein) PG], pectin methylesterase [(15.61 U mg−1 protein) PME] and β-galactosidase [(20.64 U mg−1 protein) β-Gal] enzymes. The activity of catalase [(41.56 U mg−1 protein) CAT], ascorbate peroxidase [(55.57 U mg−1 protein) APX], superoxide dismutase [(52.93 U mg−1 protein) SOD] and peroxidase [(31.02 U mg−1 protein) POD] was also higher in combined treatment. In addition, AG + GABA treated fruits showed better biochemical and sensory attributes than the control. So, combined AG + GABA could be used for CI mitigation and storage life prolongation of ‘Kinnow’ fruits.

Physiological effects of γ-aminobutyric acid application on cold tolerance in Medicago ruthenica.

Low temperatures in the seedling stage during early spring limit Medicago ruthenica germination and seedling growth. Elucidating the physiological mechanism of γ-aminobutyric acid (GABA)-regulated cold tolerance in M. ruthenica could provide a reference for alleviating the harmful effects of low temperatures on legumes in alpine meadows. The regulatory effects of GABA on M. ruthenica physiological parameters were explored by simulating the ground temperatures in the alpine meadow area of Tianzhu, China, in early May (2h at 7°C; 6h at 15°C; 4h at 12°C; 2h at 7°C; 10h at 3°C). Our results showed that 15 mmol/l GABA was the optimal spray concentration to promote growth in the aboveground and belowground parts and increase the fresh and dry weights of seedlings. At this concentration, GABA enhanced the activities of catalase, peroxidase, superoxide dismutase, and ascorbate peroxidase; increased the osmotic balance; and inhibited the production of harmful substances in the cells under low-temperature conditions. GABA also regulated the tissue structure of leaves, increased the cell tense ratio, maintained photochemical activity, increased the amount of light energy to the photochemical reaction center, and improved the photosynthetic rate. Furthermore, exogenous GABA application increased the endogenous GABA content by promoting GABA synthesis in the early stages of low-temperature stress but mainly participated in low-temperature stress mitigation via GABA degradation in the late stages. Our results show that GABA can improve the cold tolerance of M. ruthenica by promoting endogenous GABA metabolism, protecting the membrane system, and improving the leaf structure.

Polyamines Metabolism Interacts with γ-Aminobutyric Acid, Proline and Nitrogen Metabolisms to Affect Drought Tolerance of Creeping Bentgrass.

Due to increased global warming and climate change, drought has become a serious threat to horticultural crop cultivation and management. The purpose of this study was to investigate the effect of spermine (Spm) pretreatment on metabolic alterations of polyamine (PAs), γ-aminobutyric acid (GABA), proline (Pro), and nitrogen associated with drought tolerance in creeping bentgrass (Agrostis stolonifera). The results showed that drought tolerance of creeping bentgrass could be significantly improved by the Spm pretreatment, as demonstrated by the maintenance of less chlorophyll loss and higher photosynthesis, gas exchange, water use efficiency, and cell membrane stability. The Spm pretreatment further increased drought-induced accumulation of endogenous PAs, putrescine, spermidine, and Spm, and also enhanced PAs metabolism through improving arginine decarboxylases, ornithine decarboxylase, S-adenosylmethionine decarboxylase, and polyamine oxidase activities during drought stress. In addition, the Spm application not only significantly improved endogenous GABA content, glutamate content, activities of glutamate decarboxylase and α-ketoglutarase, but also alleviated decline in nitrite nitrogen content, nitrate reductase, glutamine synthetase, glutamate synthetase, and GABA aminotransferase activities under drought stress. The Spm-pretreated creeping bentgrass exhibited significantly lower ammonia nitrogen content and nitrite reductase activity as well as higher glutamate dehydrogenase activity than non-pretreated plants in response to drought stress. These results indicated beneficial roles of the Spm on regulating GABA and nitrogen metabolism contributing towards better maintenance of Tricarboxylic acid (TCA) cycle in creeping bentgrass. Interestingly, the Spm-enhanced Pro metabolism rather than more Pro accumulation could be the key regulatory mechanism for drought tolerance in creeping bentgrass. Current findings provide a comprehensive understanding of PAs interaction with other metabolic pathways to regulate drought tolerance in grass species.

Not Just a Cycle: Three gab Genes Enable the Non-Cyclic Flux Toward Succinate via GABA Shunt in 'Candidatus Liberibacter asiaticus'-Infected Citrus.

Although the mitochondria retain all required enzymes for an intact tricarboxylic acid (TCA) cycle, plants might shift the cyclic flux from the TCA cycle to an alternative noncyclic pathway via γ-aminobutyric acid (GABA) shunt under specific physiological conditions. We hypothesize that several genes may ease this noncyclic flux and contribute to the citrus response to the phytopathogenic bacterium 'Candidatus Liberibacter asiaticus', the causal agent of Huanglongbing in citrus. To test this hypothesis, we used multiomics techniques (metabolomics, fluxomics, and transcriptomics) to investigate the potential roles of putative gab homologies from Valencia sweet orange (Citrus sinensis). Our findings showed that 'Ca. L. asiaticus' significantly increased the endogenous GABA and succinate content but decreased ketoglutarate in infected citrus plants. Citrus genome harbors three putative gab genes, including amino-acid permease (also known as GABA permease; CsgabP), GABA transaminase (CsgabT), and succinate-semialdehyde dehydrogenase (also known as GABA dehydrogenase; CsgabD). The transcript levels of CsgabP, CsgabT, and CsgabD were upregulated in citrus leaves upon the infection with 'Ca. L. asiaticus' and after the exogenous application of GABA or deuterium-labeled GABA isotope (GABA-D6). Moreover, our finding showed that exogenously applied GABA is quickly converted to succinate and fed into the TCA cycle. Likewise, the fluxomics study showed that GABA-D6 is rapidly metabolized to succinate-D4. Our work proved that GABA shunt and three predicated gab genes from citrus, support the upstream noncyclic flux toward succinate rather than an intact TCA cycle and contribute to citrus defense responses to 'Ca. L. asiaticus'.[Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Regulation of nutrient accumulation by γ-aminobutyric acid associated with GABA priming-enhanced heat tolerance in creeping bentgrass

<italic>γ</italic>-Aminobutyric acid (GABA) is known for its positive effects on improving plant stress tolerance, while the association of its role in regulating nutritional availability and GABA priming-enhanced heat tolerance is not well documented. The objective of this study was to determine whether GABA priming may improve heat tolerance in cool-season grass species involving regulation of plant nutrition for macronutrients and micronutrient elements. Plants of creeping bentgrass (<italic>Agrostis stolonifera</italic>) (cv. 'Penncross') in each pot were treated with 10 mL of water (control) or 0.5 mM GABA (GABA priming) by foliar spray and then subjected to heat stress (35/30 °C, day/night) or optimal growth temperature (control) (21/19 °C, day/night) (non-stress control) for 30 d in growth chambers. GABA-primed plants had significantly higher endogenous GABA content associated with improved heat tolerance compared to non-treated plants, as reflected by higher leaf cell membrane stability, relative water content, osmotic adjustment, chlorophyll content, photochemical efficiency, and net photosynthetic rate. Plants pretreated with GABA exhibited significantly higher content of nitrogen (N), phosphorus (P), calcium (Ca), sodium (Na), and copper (Cu) but lower content of boron (B) and manganese (Mn) in leaves than non-treated plants under heat stress. The enhanced accumulation of N, P, Ca, Na, and Cu and restricted B and Mn accumulation by GABA priming indicate that GABA could modulate mineral nutrient availability in plants, contributing to improved heat tolerance for creeping bentgrass.

γ-Aminobutyric Acid (GABA) Priming Improves Seed Germination and Seedling Stress Tolerance Associated With Enhanced Antioxidant Metabolism, DREB Expression, and Dehydrin Accumulation in White Clover Under Water Stress.

As an important plant growth regulator, the role of γ-aminobutyric acid (GABA) in regulating seeds germination was less well elucidated under water stress. The present study was conducted to investigate the impact of GABA pretreatment on seeds germination of white clover (Trifolium repens) under water deficient condition. Results demonstrated that seeds pretreated with 2μmol/l GABA significantly alleviated decreases in endogenous GABA content, germination percentage, germination potential, germination index, root length, and fresh weight along with marked reduction in mean germination time after 7days of germination under drought stress. In addition, seeds priming with GABA significantly increased the accumulation of soluble sugars, non-enzymatic antioxidants [reduced ascorbate, dehydroascorbic acid, oxidized glutathione (GSSG), and reduced glutathione (GSH)], and enzymes [superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathioe reductase, and monodehydroasorbate reductase (MDHR)] activities involved in antioxidant metabolism, which could be associated with significant reduction in osmotic potential and the accumulation of superoxide anion, hydrogen peroxide, electrical leakage, and malondialdehyde in seeds under drought stress. The GABA-pretreated seeds exhibited significantly higher abundance of dehydrin (DHN, 56 KDa) and expression levels of DHNs encoding genes (SK2, Y2K, Y2SK, and Dehydrin b) and transcription factors (DREB2, DREB3, DREB4, and DREB5) than the untreated seeds during germination under water-limited condition. These results indicated that the GABA regulated improvement in seeds germination associated with enhancement in osmotic adjustment, antioxidant metabolism, and DREB-related DHNs expression. Current study will provide a better insight about the GABA-regulated defense mechanism during seeds germination under water-limited condition.

Postharvest γ-aminobutyric acid application mitigates chilling injury of aonla (Emblica officinalis Gaertn.) fruit during low temperature storage

Aonla is a nutrients rich underutilized subtropical fruit. It has short shelf life and is prone to chilling injury (CI) below 6 °C. So, the effect of γ-aminobutyric acid [5 mmol L−1 (GABA)] was investigated on aonla fruit CI mitigation during low temperature storage at 5 ± 1 °C for 24 d. Results showed that GABA treatment markedly suppressed CI, decay incidence, membrane permeability, malondialdehyde, superoxide anion and hydrogen peroxide in contrast with control. Treated fruit showed higher activity of glutamate decarboxylase (GAD) and GABA transaminase (GABA-T) enzymes which ultimately resulted in increased endogenous GABA content accumulation. Similarly, GABA application stimulated the activities of Δ1-pyrroline-5-carboxylate synthetase (P5CS) and ornithine aminotransferase (OAT) enzymes which thus enhanced proline accumulation. The GABA treated fruit exhibited higher phenylalanine ammonia lyase (PAL) and reduced polyphenol oxidase (PPO) enzyme activity and showed higher total phenols accumulation. In addition, total flavonoids, ascorbic acid and glutathione content were also significantly higher in GABA treated aonla fruit. In addition, GABA treated aonla fruit exhibited markedly higher ascorbate peroxidase (APX) catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) activities in comparison with the control. In addition, GABA treated fruit showed lower total soluble solids, ripening index and juice pH along with markedly higher concentration of titratable acidity. In conclusion, GABA treatment could be used for CI mitigation and overall quality maintenance of cold stored aonla fruit.

Alleviation of Postharvest Chilling Injury of Carambola Fruit by γ-aminobutyric Acid: Physiological, Biochemical, and Structural Characterization.

Chilling injury is a physiological disorder affecting the quality of carambola fruit. In the present study, the effect of exogenous γ-aminobutyric acid (GABA) on CI development in carambola fruit during storage at 4°C for 15 days was investigated. The results showed that 2.5-mM GABA reduced CI index, maintained pericarp lightness, and decreased the electrolyte leakage (EL) and malondialdehyde content (MDA) while increased the superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) enzyme activities. Endogenous GABA content was significantly higher in the treated fruit than in the control fruit during the whole storage. Besides, the treatment promoted the accumulation of proline and ascorbic acid (AsA) under chilling stress. Compared to the control, GABA-treated fruit exhibited a higher activity of phenylalanine ammonia-lyase (PAL) and total phenolic compounds, and a lower activity of polyphenol oxidase (PPO). In addition, the Safranin O/fast green staining revealed via microscopic images that the GABA treatment reduced the cell walls degradation of carambola fruit. Moreover, the results displayed a lower activity of phospholipase D (PLD) and lipoxygenase (LOX) enzymes, which coincided with a higher content of oleic acid (C18:1), linoleic acid (C18:2n6), and α-linolenic acid (C18:3n3) after 15 days of treatment, leading to the maintenance of the integrity and prevention of the membrane of the rapid softening of carambola fruit. The findings of the present work showed particularly new insights into the crosstalk between GABA and fatty acids. GABA might preserve the pericarp of carambola fruit by increasing the content of the unsaturated fatty acid (UFA) γ-linolenic acid and reducing the saturated fatty acid (SFA) such as caproic acid (C6:0), caprylic acid (C8:0), myristic acid (C14:0), and palmitic acid (C16:0) progressively. GABA can be used as an appropriate postharvest technology for improving the quality of carambola fruit during low-temperature storage.

Adaptability to High Temperature and Stay-Green Genotypes Associated With Variations in Antioxidant, Chlorophyll Metabolism, and γ-Aminobutyric Acid Accumulation in Creeping Bentgrass Species.

High temperature limits the cultivation and utilization of cool-season plants in many regions worldwide. Recently, extreme hot waves swept across the globe in summer, leading to enormous economic loss. The evaluation and identification of genotypic variation in thermotolerance within species are critical to breeding for environmental adaptation and also provide potential materials to explore thermo-resistant mechanism in plants. Forty-two accessions of creeping bentgrass (Agrostis stolonifera), which is a cool-season perennial grass for turf and ecological remediation, were collected from 15 different countries. Physiological traits, namely, chlorophyll (Chl) content, electrolyte leakage, photochemical efficiency, performance index on absorption basis, leaf relative water content, and osmotic potential were used to evaluate the heat tolerance of these materials in controlled growth chambers and field during summer. Stay-green and early-aging genotypes were selected to further reveal the potential mechanism of tolerance to senescence and heat damage associated with alterations in Chl metabolism, antioxidant and photosynthetic capacity, and endogenous γ-aminobutyric acid (GABA). Findings showed that there were significant genetic variations in physiological traits among 41 materials in response to high temperature stress. The 13M, PROVIDENCE, and LOFTS L-93 were the top three accessions with superior tolerance to heat and summer stress than other materials in terms of laboratory and field tests. In response to heat stress, the stay-green genotype PROVIDENCE exhibited significantly higher photochemical efficiency, net photosynthetic rate, transpiration rate, and water use efficiency than the heat-susceptible W6 6570. Delayed leaf senescence in relation to less Chl loss was detected in the PROVIDENCE associated with maintenance of significantly higher expression levels of Chl-anabolic genes (AsCHLH, AsPBGD, and AsPOR) and lower Chl-catabolic gene AsPPH under heat stress. Genetic attributes, such as better capacity to scavenge reactive oxygen species and higher endogenous GABA content could play positive roles in alleviating heat-induced senescence, oxidative damage, and metabolic disturbance in the PROVIDENCE.

Transcriptional regulation and stress-defensive key genes induced by γ-aminobutyric acid in association with tolerance to water stress in creeping bentgrass

ABSTRACT γ-Aminobutyric acid (GABA) acts as an important regulator involved in the mediation of cell signal transduction and stress tolerance in plants. However, the function of GABA in transcriptional regulation is not fully understood in plants under water stress. The creeping bentgrass (Agrostis stolonifera) was pretreated with or without GABA (0.5 mM) for 24 hours before being exposed to 5 days of water stress. Physiological analysis showed that GABA-treated plants maintained significantly higher endogenous GABA content, leaf relative water content, net photosynthetic rate, and lower osmotic potential than untreated plants under water stress. The GABA application also significantly alleviated stress-induced increases in superoxide anion (O2 .-) content, hydrogen peroxide (H2O2) content, and electrolyte leakage through enhancing total antioxidant capacity, superoxide dismutase (SOD) activity, and peroxidase (POD) activity in response to water stress. The transcriptomic analysis demonstrated that the GABA-induced changes in differentially expressed genes (DEGs) involved in carbohydrates, amino acids, and secondary metabolism helped to maintain better osmotic adjustment, energy supply, and metabolic homeostasis when creeping bentgrass suffers from water stress. The GABA triggered Ca2+-dependent protein kinase (CDPK) signaling and improved transcript levels of DREB1/2 and WRKY1/24/41 that could be associated with the upregulation of stress-related functional genes such as POD, DHNs, and HSP70 largely contributing to improved tolerance to water stress in relation to the antioxidant, prevention of cell dehydration, and protein protection in leaves.

Exogenous application of GABA retards cap browning in Agaricus bisporus and its possible mechanism

In this study, the effect of exogenous γ-aminobutyric acid (GABA) at 0 (control), 0.01, 0.1, 1, and 10 mM on cap browning and quality of button mushrooms during 15 d of storage at 4 °C was investigated. GABA at 0.1 mM yielded the lowest cap browning. Weight loss, electrolyte leakage, and malondialdehyde (MDA) content were significantly lower in mushrooms treated with 0.1 mM GABA, while their firmness was higher. The increased accumulation of total phenolic compounds in the 0.1 mM GABA treated mushrooms was associated with a higher expression and activity of phenylalanine ammonia-lyase (PAL) accompanied by a lower expression and activity of polyphenol oxidase (PPO). GABA treatment was capable of reducing H2O2 content and increasing ascorbic acid (AsA) content. Exogenous GABA increased the endogenous GABA content by increasing the expression of the glutamate decarboxylase (GAD) gene and decreasing the expression of the GABA transaminase (GABA-T) gene. Overall, GABA treatment could be employed for retarding cap browning and maintaining the sensory and nutritional quality of button mushrooms during cold storage.

γ-Aminobutyric Acid Suppresses Iron Transportation from Roots to Shoots in Rice Seedlings by Inducing Aerenchyma Formation.

γ-Aminobutyric acid (GABA) is a widely distributed non-protein amino acid mediated the regulation of nitrate uptake and Al3+ tolerance in plants. However, there are few reports about the involvement of GABA in the regulation of iron (Fe) acquisition and translocation. Here, we show that GABA regulates Fe homeostasis in rice seedlings. Exogenous GABA decreased the chlorophyll concentration in leaves, with or without Fe supply. Over-expression of glutamate decarboxylase (GAD) gene, coding a crucial enzyme of GABA production, elevated endogenous GABA content and caused more leaf chlorosis than wild type (Nipponbare). GABA inhibited Fe transportation from roots to shoots and GABA application elevated the expression levels of Fe deficiency (FD)-related genes under conditions of Fe-sufficiency (FS), suggesting that GABA is a regulator of Fe translocation. Using Perls’ blue staining, we found that more ferric iron (Fe3+) was deposited in the epidermal cells of roots treated with GABA compared with control roots. Anatomic section analysis showed that GABA treatment induced more aerenchyma formation compared with the control. Aerenchyma facilitated the oxidization of soluble ferrous iron (Fe2+) into insoluble Fe3+, resulted in Fe precipitation in the epidermis, and inhibited the transportation of Fe from roots to shoots.

Nitric Oxide Signal, Nitrogen Metabolism, and Water Balance Affected by γ-Aminobutyric Acid (GABA) in Relation to Enhanced Tolerance to Water Stress in Creeping Bentgrass.

γ-Aminobutyric acid (GABA) plays an important role in regulating stress tolerance in plants. Purposes of this study was to determine the effect of an exogenous supply of GABA on tolerance to water stress in creeping bentgrass (Agrostis stolonifera), and further reveal the GABA-induced key mechanisms related to water balance, nitrogen (N) metabolism and nitric oxide (NO) production in response to water stress. Plants were pretreated with or without 0.5 mM GABA solution in the roots for 3 days, and then subjected to water stress induced by −0.52 MPa polyethylene glycol 6000 for 12 days. The results showed that water stress caused leaf water deficit, chlorophyll (Chl) loss, oxidative damage (increases in superoxide anion, hydrogen peroxide, malondialdehyde, and protein carbonyl content), N insufficiency, and metabolic disturbance. However, the exogenous addition of GABA significantly increased endogenous GABA content, osmotic adjustment and antioxidant enzyme activities (superoxide dismutase, catalase, dehydroascorbate reductase, glutathione reductase and monodehydroascorbate reductase), followed by effectively alleviating water stress damage, including declines in oxidative damage, photoinhibition, and water and Chl loss. GABA supply not only provided more available N, but also affected N metabolism through activating nitrite reductase and glutamine synthetase activities under water stress. The supply of GABA did not increase glutamate content and glutamate decarboxylase activity, but enhanced glutamate dehydrogenase activity, which might indicate that GABA promoted the conversion and utilization of glutamate for maintaining Chl synthesis and tricarboxylic acid cycle when creeping bentgrass underwent water stress. In addition, GABA-induced NO production, depending on nitrate reductase and NO-associated protein pathways, could be associated with the enhancement of antioxidant defense. Current findings reveal the critical role of GABA in regulating signal transduction and metabolic homeostasis in plants under water-limited condition.

γ-Aminobutyric Acid Enhances Heat Tolerance Associated with the Change of Proteomic Profiling in Creeping Bentgrass.

γ-Aminobutyric acid (GABA) participates in the regulation of adaptability to abiotic stress in plants. The objectives of this study were to investigate the effects of GABA priming on improving thermotolerance in creeping bentgrass (Agrostis stolonifera) based on analyses of physiology and proteome using iTRAQ technology. GABA-treated plants maintained significantly higher endogenous GABA content, photochemical efficiency, performance index on absorption basis, membrane stability, and osmotic adjustment (OA) than untreated plants during a prolonged period of heat stress (18 days), which indicated beneficial effects of GABA on alleviating heat damage. Protein profiles showed that plants were able to regulate some common metabolic processes including porphyrin and chlorophyll metabolism, glutathione metabolism, pyruvate metabolism, carbon fixation, and amino acid metabolism for heat acclimation. It is noteworthy that the GABA application particularly regulated arachidonic acid metabolism and phenylpropanoid biosynthesis related to better thermotolerance. In response to heat stress, the GABA priming significantly increased the abundances of Cu/ZnSOD and APX4 that were consistent with superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities. The GABA-upregulated proteins in relation to antioxidant defense (Cu/ZnSOD and APX4) for the reactive oxygen species scavenging, heat shock response (HSP90, HSP70, and HSP16.9) for preventing denatured proteins aggregation, stabilizing abnormal proteins, promoting protein maturation and assembly, sugars, and amino acids metabolism (PFK5, ATP-dependent 6-phosphofructokinase 5; FK2, fructokinase 2; BFRUCT, β-fructofuranosidase; RFS2, galactinol-sucrose galactosyltransferase 2; ASN2, asparagine synthetase 2) for OA and energy metabolism, and transcription factor (C2H2 ZNF, C2H2 zinc-finger protein) for the activation of stress-defensive genes could play vital roles in establishing thermotolerance. Current findings provide an illuminating insight into the new function of GABA on enhancing adaptability to heat stress in plants.

Calcium Treatment Alleviates Pericarp Browning of 'Nanguo' Pears by Regulating the GABA Shunt After Cold Storage.

Long-term storage of pear fruit at low temperature can retard senescence but may result in pericarp browning. We previously reported that increasing endogenous γ-aminobutyrate (GABA) content by exogenous GABA treatment can maintain mitochondrial structure integrity, thereby alleviating pericarp browning of ‘Nanguo’ pears after cold storage. Here, we tested the effectiveness of Ca2+ treatment on pericarp browning in relation to GABA biosynthesis. Fruit browning was reduced by treatment with Ca2+ after 180 days of storage. Pericarp Ca2+ and calmodulin content in treated fruit increased, and concomitantly, endogenous GABA content, key GABA synthesis-related enzyme activity, and gene expression were upregulated. Moreover, the mitochondrial structure in the pericarp tissue was found to be well preserved. Thus, Ca2+ treatment effectively reduced pericarp browning of refrigerated ‘Nanguo’ pears owing to improvement in the GABA biosynthesis capacity in the fruit.

Metabolic regulation of polyamines and γ-aminobutyric acid in relation to spermidine-induced heat tolerance in white clover.

Heat stress decreases crop growth and yield worldwide. Spermidine (Spd) is a small aliphatic amine and acts as a ubiquitous regulator for plant growth, development and stress tolerance. Objectives of this study were to determine effects of exogenous Spd on changes in endogenous polyamine (PA) and γ-aminobutyric acid (GABA) metabolism, oxidative damage, senescence and heat shock protein (HSP) expression in white clover subjected to heat stress. Physiological and molecular methods, including colorimetric assay, high performance liquid chromatography and qRT-PCR, were applied. Results showed that exogenous Spd significantly alleviated heat-induced stress damage. Application of Spd not only increased endogenous putrescine, Spd, spermine and total PA accumulation, but also accelerated PA oxidation and improved glutamic acid decarboxylase activity, leading to GABA accumulation in leaves under heat stress. The Spd-pretreated white clover maintained a significantly higher chlorophyll (Chl) content than untreated plants under heat stress, which could be related to the roles of Spd in up-regulating genes encoding Chl synthesis (PBGD and Mg-CHT) and maintaining reduced Chl degradation (PaO and CHLASE) during heat stress. In addition, Spd up-regulated HSP70, HSP70B and HSP70-5 expression, which might function in stabilizing denatured proteins and helping proteins to folding correctly in white clover under high temperature stress. In summary, exogenous Spd treatment improves the heat tolerance of white clover by altering endogenous PA and GABA content and metabolism, enhancing the antioxidant system and HSP expression and slowing leaf senescence related to an increase in Chl biosynthesis and a decrease in Chl degradation during heat stress.

Effects of GABA and Vigabatrin on the Germination of Chinese Chestnut Recalcitrant Seeds and Its Implications for Seed Dormancy and Storage.

Recalcitrant chestnut seeds are rich in γ-aminobutyric acid (GABA), which negatively regulates adventitious root development by altering carbon/nitrogen metabolism. However, little is known regarding the role of this metabolite in chestnut seeds. In this study, we investigated the effects of GABA changes on the germination of chestnut seeds treated with exogenous GABA and vigabatrin (VGB, which inhibits GABA degradation). Both treatments significantly inhibited seed germination and primary root growth and resulted in the considerable accumulation of H2O2, but the endogenous GABA content decreased before germination at 48 h. Soluble sugar levels increased before germination, but subsequently decreased, whereas starch contents were relatively unchanged. Changes to organic acids were observed at 120 h after sowing, including a decrease and increase in citrate and malate levels, respectively. Similarly, soluble protein contents increased at 120 h, but the abundance of most free amino acids decreased at 48 h. Moreover, the total amino acid levels increased only in response to VGB at 0 h. Accordingly, GABA and VGB altered the balance of carbon and nitrogen metabolism, thereby inhibiting chestnut seed germination. These results suggested that changes to GABA levels in chestnut seeds might prevent seed germination. The study data may also help clarify the dormancy and storage of chestnut seeds, as well as other recalcitrant seeds.

Regulation of Heat Shock Factor Pathways by γ-aminobutyric Acid (GABA) Associated with Thermotolerance of Creeping Bentgrass.

Activation and enhancement of heat shock factor (HSF) pathways are important adaptive responses to heat stress in plants. The γ-aminobutyric acid (GABA) plays an important role in regulating heat tolerance, but it is unclear whether GABA-induced thermotolerance is associated with activation of HSF pathways in plants. In this study, the changes of endogenous GABA level affecting physiological responses and genes involved in HSF pathways were investigated in creeping bentgrass during heat stress. The increase in endogenous GABA content induced by exogenous application of GABA effectively alleviated heat damage, as reflected by higher leaf relative water content, cell membrane stability, photosynthesis, and lower oxidative damage. Contrarily, the inhibition of GABA accumulation by the application of GABA biosynthesis inhibitor further aggravated heat damage. Transcriptional analyses showed that exogenous GABA could significantly upregulate transcript levels of genes encoding heat shock factor HSFs (HSFA-6a, HSFA-2c, and HSFB-2b), heat shock proteins (HSP17.8, HSP26.7, HSP70, and HSP90.1-b1), and ascorbate peroxidase 3 (APX3), whereas the inhibition of GABA biosynthesis depressed these genes expression under heat stress. Our results indicate GABA regulates thermotolerance associated with activation and enhancement of HSF pathways in creeping bentgrass.