Proline Accumulation Research Articles

RBOH-dependent signaling is involved in He-Ne laser-induced salt tolerance and production of rosmarinic acid and carnosol in Salvia officinalis

BackgroundIn the past two decades, the impacts of Helium-Neon (He–Ne) laser on stress resistance and secondary metabolism in plants have been studied, but the signaling pathway which by laser regulates this process remains unclear. Therefore, the current study sought to explore the role of RBOH-dependent signaling in He–Ne laser-induced salt tolerance and elicitation of secondary metabolism in Salvia officinalis. Seeds were primed with He–Ne laser (6 J cm− 2) and peroxide hydrogen (H2O2, 5 mM) and 15-old-day plants were exposed to two salinity levels (0, 75 mM NaCl).ResultsSalt stress reduced growth parameters, chlorophyll content and relative water content (RWC) and increased malodialdehyde (MDA) and H2O2 contents in leaves of 45-old-day plants. After 48 h of salt exposure, higher transcription levels of RBOH (encoding NADPH oxidase), PAL (phenylalanine ammonia-lyase), and RAS (rosmarinic acid synthase) were recorded in leaves of plants grown from seeds primed with He–Ne laser and/or H2O2. Despite laser up-regulated RBOH gene in the early hours of exposing to salinity, H2O2 and MDA contents were lower in leaves of these plants after 30 days. Seed pretreatment with He–Ne laser and/or H2O2 augmented the accumulation of anthocyanins, total phenol, carnasol, and rosmarinic acid and increased total antioxidant capacity under non-saline and more extensively at saline conditions. Indeed, these treatments improved RWC, and K+/Na+ ratio, enhanced the activities of superoxide dismutase and ascorbate peroxidase and proline accumulation, and significantly decreased membrane injury and H2O2 content in leaves of 45-old-day plants under salt stress. However, applying diphenylene iodonium (DPI as an inhibitor of NADPH oxidase) and N, N-dimethyl thiourea (DMTU as a H2O2 scavenger) after laser priming reversed the aforementioned effects which in turn resulted in the loss of laser-induced salt tolerance and secondary metabolism.ConclusionsThese findings for the first time deciphered that laser can induce a transient RBOH-dependent H2O2 burst, which might act as a downstream signal to promote secondary metabolism and salt stress alleviation in S. officinalis plants.

WRKY transcription factor 40 from eggplant (Solanum melongena L.) regulates ABA and salt stress responses

Plants are affected by many environmental factors during their various stages of growth, among which salt stress is a key factor. WRKY transcription factors play important roles in the response to stress in plants. In this study, SmWRKY40 from eggplant (Solanum melongena L.) was found to belong to the subfamily of WRKY transcription factor group II, closely related to the evolution of wild tomato ScWRKY40 (Solanum chilense). The expression of SmWRKY40 could be induced by several abiotic stresses (drought, salt, and high temperature) and ABA to different degrees, with salt stress being the most significant. In Arabidopsis thaliana, the seed germination rate of SmWRKY40 overexpression seedlings was significantly higher than those of the wild type under high concentrations of NaCl and ABA, and root elongation of overexpression lines was also longer than wild type under NaCl treatments. SmWRKY40 overexpression lines were found to enhance Arabidopsis tolerance to salt with lower ROS, MDA, higher soluble protein, proline accumulation, and more active antioxidant enzymes. The expression level of genes related to stress and ABA signaling displayed significant differences in SmWRKY40 overexpression line than that of WT. These results indicate that SmWRKY40 regulates ABA and salt stress responses in Arabidopsis.

SlWRKY51 regulates proline content to enhance chilling tolerance in tomato.

Chilling stress is a major environmental factor that significantly reduces crop production. To adapt to chilling stress, plants activate a series of cellular responses and accumulate an array of metabolites, particularly proline. Here, we report that the transcription factor SlWRKY51 increases proline contents in tomato (Solanum lycopersicum) under chilling stress. SlWRKY51 expression is induced under chilling stress. Knockdown or knockout of SlWRKY51 led to chilling-sensitive phenotypes, with lower photosynthetic capacity and more reactive oxygen species (ROS) accumulation than the wild type (WT). The proline contents were significantly reduced in SlWRKY51 knockdown and knockout lines under chilling stress, perhaps explaining the phenotypes of these lines. D-1-pyrroline-5-carboxylate synthetase (P5CS), which catalyses the rate-limiting step of proline biosynthesis, is encoded by two closely related P5CS genes (P5CS1 and P5CS2). We demonstrate that SlWRKY51 directly activates the expression of P5CS1 under chilling stress. In addition, the VQ (a class of plant-specific proteins containing the conserved motif FxxhVQxhTG) family member SlVQ10 physically interacts with SlWRKY51 to enhance its activation of P5CS1. Our study reveals that the chilling-induced transcription factor SlWRKY51 enhances chilling tolerance in tomato by promoting proline accumulation.

The nonphototrophic hypocotyl 3 (NPH3) domain protein NRL5 is a trafficking-associated GTPase essential for drought resistance.

The mechanisms of plant drought resistance are unclear but may involve membrane trafficking and metabolic reprogramming, including proline accumulation. Forward genetic screening using a proline dehydrogenase 1 (ProDH1) promoter:reporter identified a drought hypersensitive mutant with a single-amino acid substitution (P335L) in the nonphototrophic hypocotyl 3 (NPH3) domain of NPH3/root phototropism 2-like 5 (NRL5)/naked pins in Yucca 8 (NPY8). Further experiments found that NRL5 and other NPH3 domain proteins are guanosine triphosphatases (GTPases). NRL5, but not NRL5P335L, interacted with the RABE1c and RABH1b GTPases and the soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) Vesicle-Associated Membrane Protein (VAMP)721/722. These proteins controlled NRL5 localization and connection to trafficking while also being genetically downstream of, and potentially regulated by, NRL5. These data demonstrate that NRL5-mediated restraint of proline catabolism is required for drought resistance and also reveal unexpected functions of the NPH3 domain such that the role of NPH3 domain proteins in signaling, trafficking, and cellular polarity can be critically reevaluated.

Overexpression of Potato PYL16 Gene in Tobacco Enhances the Transgenic Plant Tolerance to Drought Stress.

PYR/PYL/RCAR proteins are abscisic acid (ABA) receptors that play a crucial role in plant responses to abiotic stresses. However, there have been no research reports on potato PYL so far. In this study, a potato PYL gene named StPYL16 was identified based on transcriptome data under drought stress. Molecular characteristics analysis revealed that the StPYL16 protein possesses an extremely conserved PYL family domain. The tissue expression results indicated that the StPYL16 is predominantly expressed at high levels in the underground parts, particularly in tubers. Abiotic stress response showed that StPYL16 has a significant response to drought treatment. Further research on the promoter showed that drought stress could enhance the activation activity of the StPYL16 promoter on the reporter gene. Then, transient and stable expression of StPYL16 in tobacco enhanced the drought resistance of transgenic plants, resulting in improved plant height, stem thickness, and root development. In addition, compared with wild-type plants, StPYL16 transgenic tobacco exhibited lower malondialdehyde (MDA) content, higher proline accumulation, and stronger superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities. Meanwhile, StPYL16 also up-regulated the expression levels of stress-related genes (NtSOD, NtCAT, NtPOD, NtRD29A, NtLEA5, and NtP5CS) in transgenic plants under drought treatment. These findings indicated that the StPYL16 gene plays a positive regulatory role in potato responses to drought stress.

Modulatory effects of selenium nanoparticles against drought stress in some grapevine rootstock/scion combinations

BackgroundDrought is a significant abiotic stress that adversely affects plant growth, development, and metabolic processes, thereby reducing plant yield, quality, and production, and threatening global food security. In recent years, nanotechnology has emerged as a promising strategy to overcome the existing environmental challenges and has been tested on some plant species. But it is still awaiting investigation for grapevines. The aim of this study was to investigate the potential of selenium nanoparticles (Se-NPs) to modulate some morphological, physiological, and biochemical parameters in grapevine saplings (5 BB/Crimson Seedless, 41 B/Crimson Seedless, and 1103 P/Crimson Seedless) under drought stress conditions.ResultsIn the study, Se-NP solutions at different concentrations (0 (control), 1, 10, and 100 ppm) were applied by the spray method to wet the entire green surface of grapevine saplings grown under well-irrigated (90–100% field capacity) and drought stress (40–50% field capacity) conditions. Our results showed that 10 ppm Se-NP concentration had the most positive effect, 1 ppm concentration showed limited effects, and 100 ppm concentration led to toxic effects, especially when combined with drought conditions. Se-NP applications at 10 ppm concentration improved the growth parameters (leaf number, leaf area, root fresh and dry weight, shoot fresh and dry weight, etc.) and increased the SPAD index of grapevine saplings under both normal and drought conditions. Additionally, 10 ppm Se-NP applications improved the relative water content (RWC) and stomatal conductance values, proportional to the increases in protein content. On the other hand, under drought conditions, the drought index, leaf temperature, membrane damage index, hydrogen peroxide (H2O2) content, and malondialdehyde (MDA) levels significantly decreased as a result of 10 ppm Se-NP applications, showing an opposite trend. Furthermore, the levels of proline, total phenolics, and antioxidant enzymes (CAT, SOD, and APX) that rose significantly due to drought stress were reduced by 10 ppm Se-NP applications, which also helped to lessen the oxidative stress caused by the drought.ConclusionThe study concluded that foliar application of Se-NPs at 10 ppm significantly enhances drought tolerance in grapevine saplings by improving antioxidant defense, proline and protein accumulation, and overall growth, while lower concentrations are less effective and higher concentrations can cause phytotoxicity. These findings indicate that Se-NPs applications may hold promise not only for grapevines but also for mitigating drought stress effects and improving productivity in other economically important fruit species, warranting further exploration across diverse crop systems.Graphical

Foliar spraying with amino acids and their chitosan nanocomposites as promising way to alleviate abiotic stress in iceberg lettuce grown at different temperatures

We analyzed the effects of foliar spraying with amino acids, chitosan (CHS) and nanocomposites (NCs) of chitosan with the amino acids proline, l-cysteine and glycine betaine (CHS-Pro NCs; CHS-Cys NCs, CHS-GB NCs, respectively) on the changes in the physiological and biochemical parameters of iceberg lettuce grown at the control temperature (20 °C) and under chilling conditions (4 °C). The physicochemical parameters of the phospholipid monolayers (PLs) extracted from plants showed the effects of the treatments on the properties of the monolayers, namely, the packing density and flexibility. We observed increased accumulation of proline at 4 °C, and differences in the concentrations of sugars in most of the analyzed variants were a consequence of the lowered temperature and/or the use of organic compounds. A temperature of 4 °C caused a significant increase in the l-ascorbic acid level compared with that at 20 °C. Differences were also found in glutathione (GSH) content depending on the temperature and treatment with the tested organic compounds. CHS NCs loaded with Pro and GB were effective at increasing the amount of phenols under stress temperature conditions. We noted that a significant increase in the antioxidant activity of plants at 4 °C occurred after priming with Cys, CHS-Cys NCs, Pro and CHS-Pro NCs, and the CHS nanocomposites were more effective in this respect. Both low-temperature stress and foliar spraying of lettuce with various organic compounds caused changes in the activity of antioxidant enzymes. Two forms of dismutase (SOD), iron superoxide dismutase (FeSOD) and copper/zinc superoxide dismutase (Cu/ZnSOD), were identified in extracts from the leaves of iceberg lettuce seedlings. The application of the tested organic compounds, alone or in combination with CHS, increased the amount of malondialdehyde (MDA) in plants grown under controlled temperature conditions. Chilling caused an increase in the content of MDA, but some organic compounds mitigated the impact of low temperature. Compared with that of plants subjected to 20 °C, the fresh weight of plants exposed to chilling decreased. However, the tested compounds caused a decrease in fresh weight at 4 °C compared with the corresponding control samples. An interesting exception was the use of Cys, for which the difference in the fresh weight of plants grown at 20 °C and 4 °C was not statistically significant. After Cys application, the dry weight of the chilled plants was greater than that of the chilled control plants but was also greater than that of the other treated plants in this group. To our knowledge, this is the first report demonstrating that engineered chitosan–amino acid nanocomposites could be applied as innovative protective agents to mitigate the effects of chilling stress in crop plants.

An ecotype-specific effect of osmopriming and melatonin during salt stress in Arabidopsis thaliana

BackgroundNatural populations of Arabidopsis thaliana exhibit phenotypic variations in specific environments and growth conditions. However, this variation has not been explored after seed osmopriming treatments. The natural variation in biomass production and root system architecture (RSA) was investigated across the Arabidopsis thaliana core collection in response to the pre-sawing seed treatments by osmopriming, with and without melatonin (Mel). The goal was to identify and characterize physiologically contrasting ecotypes.ResultsVariability in RSA parameters in response to PEG-6000 seed osmopriming with and without Mel was observed across Arabidopsis thaliana ecotypes with especially positive impact of Mel addition under both control and 100 mM NaCl stress conditions. Two ecotypes, Can-0 and Kn-0, exhibited contrasted root phenotypes: seed osmopriming with and without Mel reduced the root growth of Can-0 plants while enhancing it in Kn-0 ones under both control and salt stress conditions. To understand the stress responses in these two ecotypes, main stress markers as well as physiological analyses were assessed in shoots and roots. Although the effect of Mel addition was evident in both ecotypes, its protective effect was more pronounced in Kn-0. Antioxidant enzymes were induced by osmopriming with Mel in both ecotypes, but Kn-0 was characterized by a higher responsiveness, especially in the activities of peroxidases in roots. Kn-0 plants experienced lower oxidative stress, and salt-induced ROS accumulation was reduced by osmopriming with Mel. In contrast, Can-0 exhibited lower enzyme activities but the accumulation of proline in its organs was particularly high. In both ecotypes, a greater response of antioxidant enzymes and proline accumulation was observed compared to mechanisms involving the reduction of Na+ content and prevention of K+ efflux.ConclusionsIn contrast to Can-0, Kn-0 plants grown from seeds osmoprimed with and without Mel displayed a lower root sensitivity to NaCl-induced oxidative stress. The opposite root growth patterns, enhanced by osmopriming treatments might result from different protective mechanisms employed by these two ecotypes which in turn result from adaptive strategies proper to specific habitats from which Can-0 and Kn-0 originate. The isolation of contrasting phenotypes paves the way for the identification of genetic factors affecting osmopriming efficiency.

Revitalizing maize growth and yield in water-limited environments through silicon and zinc foliar applications

Maize is an economically vital cereal crop. However, water deficiency can severely impact its productivity. Thus, it is necessary to implement an essential approach to increase maize yield while navigating the limitations imposed by scarce water supplies. The present study aimed to investigate whether foliar applications of silicon (Si) and zinc (Zn) could mitigate the adverse effects of water deficiency and improve maize growth and yield. Field experiments were conducted in Egypt during two growing seasons (2021–2022) under three irrigation regimes: full irrigation (ET0), moderate stress (ET1), and severe stress (ET2). The treatments comprised foliar sprays of Si, Zn, Si + Zn, and water control. Phenological, growth, physiological, chemical, and yield-related traits were assessed. Results showed that adequate irrigation (ET0) enhanced most parameters compared to water stress treatments. Under ET0, the combined silicon and zinc treatment resulted in the highest values for plant height, leaf area, chlorophyll content, grains per ear, kernel weight, ear size, and yield compared to other foliar treatments. Under drought stress (ET1, ET2), Si + Zn applications maintained superiority in mitigating yield losses. Proline accumulation was highest under severe stress (ET2) in the absence of foliar sprays, indicating greater drought impacts. Correlation analysis revealed positive associations of grain yield with ear size, leaf area, kernel weight, and biological yield. Cluster analysis separated irrigation regimes and visualized the consistently beneficial effects of Si + Zn across all water levels. Overall, the results demonstrate the synergistic potential of Si and Zn supplementation to sustain maize performance and yields under varying water availability.

Metal Sulfates and Microbial Food Treatment Alleviate the Oxidative Damage Caused by PEG-Induced Osmotic Stress in Cotton Plants

Background: Drought stress restricts cotton production, which causes a considerable loss of cotton yields worldwide. Plant activators improve crop production and ameliorate abiotic stresses. Thus, the effects of plant activators on the genotype of cotton (Gossypium hirsutum L.) Candia, which is resistant to drought, were investigated.Methods: Plants were treated with the plant activators, and then osmotic stress was initiated by 30% PEG 6000 treatment for 13 days. Antioxidant enzyme activities such as superoxide dismutase (SOD), ascorbate peroxidase (APX), and peroxidase (POX) were measured along with growth, relative water content (RWC), osmotic potential, photosynthetic pigments, ion leakage, proline accumulation, malondialdehyde (MDA) content, and reactive oxygen species (ROS).Results: Osmotic stress reduced shoot fresh weight (FW) and osmotic potential but increased root length of Candia. Activator treatment alleviated these effects. Photosynthetic pigment contents and cell membrane permeability were not affected by osmotic stress and activator treatments; proline accumulation and lipid peroxidation levels were enhanced by osmotic stress, but activators alleviated these increments. Activity bands for the isozymes Mn-SOD1, SOD2, and SOD3, as well as the isozyme Fe-SOD, were found in Candia. Osmotic stress elevated APX activity in Candia leaves, while activator treatment decreased it. POX activity was not affected by osmotic stress, but activator treatment decreased POX activity. Osmotic stress-induced accumulation of ROS was lessened by activator treatment.Conclusion: Plant activator treatment reduced the amount of oxidative damage (ROS buildup and lipid peroxidation) caused by drought stress in the Candia genotype.Keywords: Antioxidant system; Cotton; Gossypium hirsutum L.; Osmotic stress; PEG 6000; Plant activators

Tetragonal crystalline MnO nanoparticles alleviate Pb stress in wheat by modulating antioxidant enzymes in leaves.

Agriculture ecosystems are seriously threatened by lead (Pb) contamination, which impacts plant growth and productivity. In this study, green synthesized manganese oxide nanoparticles (MnO NPs) using citrus peel were used for priming of wheat seeds. For the synthesis of MnO nanoparticles, peel extract of Citrus paradisi and 1mM solution of manganese acetate were stirred and calcinated at 500°C. Successful synthesis of MnO NPs was determined using advanced techniques. In Fourier-transform infrared spectroscopy (FTIR), the presence of amines, alkanes, aldehydes, and alcohol molecules, on the surface of MnO NPs, confirmed their stability. X-ray diffraction analysis described their average size (22nm), while scanning electron microscopy showed tetragonal crystalline shape and nano-flowers structure of MnO NPs. Sharp peaks of energy dispersive x-ray analysis described the presence of oxygen (28.81%) and manganese (71.19%) on MnO NPs. Priming of wheat seeds with synthesized MnO NPs significantly improved the growth attributes of wheat seedlings including the size of leaf, root length, size of shoots, chlorophyll and carotenoid contents, relative water content, decreased relative electrolyte leakage, high proline accumulation and decreased concentration of malondialdehyde. Application of MnO NPs also helped plants to accumulate antioxidant enzymes in their leaves. These results proved that the priming of MnO NPs can greatly reduce lead-induced stress in wheat seedlings and these NPs can alsobe used for the priming of other crops.

ZmDST44 Gene Is a Positive Regulator in Plant Drought Stress Tolerance.

Improving drought tolerance in plants is essential for increasing crop yields under water-limited conditions. In this study, we investigated the functional role of the maize gene ZmDST44, which is targeted by the miRNA ZmmiR139. Our results indicate that ZmmiR139 regulates ZmDST44 by cleaving its mRNA, as confirmed by inverse expression patterns and 5'-RACE analysis. Overexpression of ZmDST44 in Arabidopsis, rice, and maize resulted in significant enhancements in drought tolerance. Transgenic plants exhibited reduced malondialdehyde (MDA) levels, increased proline accumulation, and upregulation of drought-responsive genes compared to wild-type plants. Transgenic Arabidopsis and rice showed improved drought resistance and higher post-drought recovery rates, and transgenic maize displayed lower sensitivity to drought stress. These findings suggest that ZmDST44 acts as a positive regulator of drought tolerance across different plant species and holds promise for developing drought-resistant crops through genetic engineering.

Impact of a biostimulant enriched in betalain degradation products on ROS signaling, proline accumulation, and phytohormone homeostasis

This study investigates the potential of a biostimulant derived from Selenicereus undatus peel waste and enriched in betalain degradation products (BDP), to influence Arabidopsis thaliana seedling development. Notably, lower BDP concentrations enhanced seedling development, while higher dosages exhibited adverse effects. Assessment of mitochondrial activity in both seeds and purified organelles showed that the tested biostimulant did not affect mitochondrial activity or integrity, highlighting its independence from mitochondrial performance. Mechanistically, BDP-enriched biostimulant modulated ROS-signaling, diminishing H2O2 by regulating the enzymatic activity and gene expression of SOD, CAT, GPX, and GR. Particularly, analyzing their different isoform via qRT-PCR, the primary cellular compartment where detoxification occurred were identified. Furthermore, biostimulant was able to influence proline-accumulation, altering both the expression of metabolism (PC5S, P5CR and OAT) and catabolism (PDH and P5CDH) related genes. Finally, the BDP-enriched biostimulant altered phytohormone levels, mainly affecting ABA/ABA-glu, tZea/tZea-rib, and tZea/IAA. Concerning GAs, the increase in GA4 and GA7 suggested an involvement of GA13ox, a hypothesis encouraged by qRT-PCR analysis. In summary, this study underscores the potential of BDP-based biostimulant as sustainable promoters of plant growth, influencing critical regulatory pathways during germination. Further research is necessary to explore their extensive applications in agricultural practices.

Jasmonic Acid and Salicylic Acid improved resistance against Spodoptera frugiperda Infestation in maize by modulating growth and regulating redox homeostasis

Exploring host plant resistance and elevating plant defense mechanisms through the application of exogenous elicitors stands as a promising strategy for integrated pest management. The fall armyworm, a pernicious menace to grain crops in tropical and subtropical regions, stands as a formidable threat due to its capacity for devastation and a wide-ranging spectrum of host plants. There is no literature regarding artificially induced resistance in maize against fall armyworm (Spodoptera frugiperda) by exogenous application of phytohormones. The present investigation was performed to evaluate the role of jasmonic acid (JA) and salicylic acid (SA) on two maize hybrids namely FH-1046 and YH-1898 against fall armyworm. Results showed that plant height, biomass and lengths, fresh and dry weight of root shoot which decreased with armyworm infestation improved with phytohormonal application. JA treatment resulted in a higher increase in all attributes as compared to SA treatment. Improvement in relative water contents, photosynthetic pigments and pronounced levels of phenol and proline accumulation were observed in infested plants after JA treatment. Infested plants recovered from oxidative stress as JA application activated and increased the antioxidant enzyme activity of superoxide dismutase, peroxidase and polyphenol oxidase activity in both FH-1046 and YH-1898 . The oxidative stress reduction in infested plants after JA treatment was also evident from a fair decrease in MDA and H2O2 in both varieties. The SA and JA mediated genes expression was studied and it was found that in FH1046 maize cultivar, JA dependent genes, particularly marker genes PR1 and Lox5 were highly expressed along with TPS10 and BBT12. Whereas SPI, WRKY28, ICS and PAL were shown to be activated upon SA application. Evidently, both JA and SA elicited a robust defensive response within the maize plants against the voracious S. frugiperda, which in consequence exerted a discernible influence over the pest's developmental trajectory and physiological dynamics. A decrease in detoxification enzyme activity of the insects was observed after feeding on treated plants. Moreover, it was recorded that the survival and weight gain of FAW feeding on phytohormone treated maize plants also decelerated. In conclusion, FH-1046 was found to be more tolerant than YH-1898 against fall armyworm infestation and 1 mM JA was more effective than 1 mM SA for alleviation of fall armyworm stress. Therefore, it was inferred that phytohormones regulated redox homeostasis to circumvent oxidative damage and mediate essential metabolic events in maize under stress. To our current understanding, this study is the very first presentation of induced resistance in maize against S. frugiperda with the phytohormonal application (JA and SA).

Enhancing tomato plants' tolerance to combined heat and salt stress – The role of arbuscular mycorrhizae and biochar

The Mediterranean basin is highly susceptible to climate change, with soil salinization and the increase in average temperatures being two of the main factors affecting crop productivity in this region. Following our previous studies on describing the detrimental effects of heat and salt stress co-exposure on tomato plants, this study aimed to understand if substrate supplementation with a combination of arbuscular mycorrhizal fungi (AMF) and biochar could mitigate the negative consequences of these stresses. Upon 21 days of exposure, stressed tomato plants grown under supplemented substrates showed increased tolerance to heat (42 °C for 4 h/day), salt (100 mM NaCl), and their combination, presenting increased biomass and flowering rate. The beneficial effects of AMF and biochar were associated with a better ionic balance (i.e. lower sodium accumulation and higher uptake of calcium and magnesium) and increased photosynthetic efficiency. Indeed, these plants presented higher chlorophyll content and improved CO2 assimilation rates. Biochemical data further supported that tomato plants grown with AMF and biochar were capable of efficiently modulating their defence pathways, evidenced by the accumulation of proline, ascorbate, and glutathione, coupled with a lower dependency on energy-costly enzymatic antioxidant players. In summary, the obtained data strongly point towards a beneficial role of combined AMF and biochar as sustainable tools to improve plant growth and development under a climate change scenario, where soil salinization and heat peaks often occur together.

Effects of High Salinity and Water Stress on Wetland Grasses from the Spanish Mediterranean Coast.

The impacts of climate change are reaching unprecedented levels, heightening the risk of species loss and ecosystem service degradation. Wetlands, highly threatened ecosystems, serve vital ecological functions by capturing carbon, filtering water, and harbouring diverse wildlife. Coastal wetlands encounter many challenges, such as increased drought periods and escalating salinity levels, severely impacting plant biodiversity. Assessing how plants respond to various environmental stress factors is imperative for devising successful conservation strategies. In the present study, we examined three representative grass species found in various habitats within the Albufera Natural Park, close to the city of Valencia on the Spanish Mediterranean coast: Imperata cylindrica, Phragmites australis, and Saccharum ravennae. High salinity and water stress conditions were induced by subjecting the plants to irrigation with solutions containing 200, 400, 600, and 800 mM NaCl or withholding irrigation altogether to mimic coastal flooding and drought scenarios. The treatments were maintained until noticeable wilting of the plants occurred, at which point a range of stress biomarkers were determined, including photosynthetic pigments, ions, osmolytes, oxidative stress markers, and antioxidant metabolites, as well as antioxidant enzyme activities. Saccharum ravennae displayed the highest sensitivity to salt stress, whereas I. cylindrica appeared to be the most tolerant. The primary salinity tolerance mechanism observed in I. cylindrica and P. australis was a blockage of ion transport from the root zone to the aerial part, together with the salt-induced accumulation of proline and soluble sugars to high concentrations in the former. No significant effects of the water deficit treatment on the growth or biochemical parameters were observed for any of the analysed species. These findings offer valuable information for the effective management and conservation of coastal wetlands facing the challenges posed by climate change.

Regulation of Proline Accumulation and Protein Secretion in Sorghum under Combined Osmotic and Heat Stress.

Plants reprogramme their proteome to alter cellular metabolism for effective stress adaptation. Intracellular proteomic responses have been extensively studied, and the extracellular matrix stands as a key hub where peptide signals are generated/processed to trigger critical adaptive signal transduction cascades inaugurated at the cell surface. Therefore, it is important to study the plant extracellular proteome to understand its role in plant development and stress response. This study examined changes in the soluble extracellular sub-proteome of sorghum cell cultures exposed to a combination of sorbitol-induced osmotic stress and heat at 40 °C. The combined stress significantly reduced metabolic activity and altered protein secretion. While cells treated with osmotic stress alone had elevated proline content, the osmoprotectant in the combined treatment remained unchanged, confirming that sorghum cells exposed to combined stress utilise adaptive processes distinct from those invoked by the single stresses applied separately. Reactive oxygen species (ROS)-metabolising proteins and proteases dominated differentially expressed proteins identified in cells subjected to combined stress. ROS-generating peroxidases were suppressed, while ROS-degrading proteins were upregulated for protection from oxidative damage. Overall, our study provides protein candidates that could be used to develop crops better suited for an increasingly hot and dry climate.

Genome-wide and functional analysis of late embryogenesis abundant (LEA) genes during dormancy and sprouting periods of kernel consumption apricots (P. armeniaca L. × P. sibirica L.)

Late embryogenesis abundant (LEA) proteins play a crucial role in protecting cells from stress, making them potential contributors to abiotic stress tolerance. This study focuses on apricot (P. armeniaca L. × P. sibirica L.), where a comprehensive genome-wide analysis identified 54 LEA genes, categorized into eight subgroups based on phylogenetic relationships. Synteny analysis revealed 14 collinear blocks containing LEA genes between P. armeniaca × P. sibirica and Arabidopsis thaliana, with an additional 9 collinear blocks identified between P. armeniaca × P. sibirica and poplar. Examination of gene structure and conserved motifs indicated that these subgroups exhibit consistent exon-intron patterns and shared motifs. The expansion and duplication of LEA genes in P. armeniaca × P. sibirica were driven by whole-genome duplication (WGD), segmental duplication, and tandem duplication events. Expression analysis, utilizing RNA-seq data and quantitative real-time RT-PCR (qRT-PCR), indicated induction of PasLEA2-20, PasLEA3-2, PasLEA6-1, Pasdehydrin-3, and Pasdehydrin-5 in flower buds during dormancy and sprouting phases. Coexpression network analysis linked LEA genes with 15 cold-resistance genes. Remarkably, during the four developmental stages of flower buds in P. armeniaca × P. sibirica - physiological dormancy, ecological dormancy, sprouting period, and germination stage - the expression patterns of all PasLEAs coexpressed with cold stress-related genes remained consistent. Protein-protein interaction networks, established using Arabidopsis orthologs, emphasized connections between PasLEA proteins and cold resistance pathways. Overexpression of certain LEA genes in yeast and Arabidopsis conferred advantages under cold stress, including increased pod length, reduced bolting time and flowering time, improved survival and seed setting rates, elevated proline accumulation, and enhanced antioxidative enzymatic activities. Furthermore, these overexpressed plants exhibited upregulation of genes related to flower development and cold resistance. The Y1H assay confirmed that PasGBF4 and PasDOF3.5 act as upstream regulatory factors by binding to the promoter region of PasLEA3-2. PasDOF2.4, PasDnaJ2, and PasAP2 were also found to bind to the promoter of Pasdehydrin-3, regulating the expression levels of downstream genes. This comprehensive study explores the evolutionary relationships among PasLEA genes, protein interactions, and functional analyses during various stages of dormancy and sprouting in P. armeniaca × P. sibirica. It offers potential targets for enhancing cold resistance and manipulating flower bud dormancy in this apricot hybrid.

Foliar Application of Chitosan (CTS), γ-Aminobutyric Acid (GABA), or Sodium Chloride (NaCl) Mitigates Summer Bentgrass Decline in the Subtropical Zone.

Creeping bentgrass (Agrostis stolonifera) is an excellent cool-season turfgrass that is widely used in urban gardening, landscaping, and golf turf. Triennial field experiments from 2017 to 2019 were conducted to investigate effects of the foliar application of chitosan (CTS), γ-aminobutyric acid (GABA), or sodium chloride (NaCl) on mitigating summer bentgrass decline (SBD) and exploring the CTS, GABA, or NaCl regulatory mechanism of tolerance to summer heat stress associated with changes in chlorophyll (Chl) loss and photosynthetic capacity, osmotic adjustment (OA), oxidative damage, and cell membrane stability. The findings demonstrated that persistent ambient high temperatures above 30 °C during the summer months of 2017, 2018, and 2019 significantly reduced the turf quality (TQ), Chl content, photochemical efficiency of PSII (Fv/Fm and PIABS), leaf relative water content, and osmotic potential (OP) but significantly increased electrolyte leakage (EL) and the accumulations of free proline, water-soluble carbohydrate (WSC), hydrogen peroxide (H2O2), and malondialdehyde (MDA). The foliar application of CTS, GABA, or NaCl could significantly alleviate SBD, as reflected by improved TQ and delayed Chl loss during hot summer months. Heat-induced declines in Fv/Fm, PIABS, the net photosynthetic rate (Pn), the transpiration rate (Tr), and water use efficiency (WUE) could be significantly mitigated by the exogenous application of CTS, GABA, or NaCl. In addition, the foliar application of CTS, GABA, or NaCl also significantly improved the accumulations of free proline and WSC but reduced the EL, OP, and H2O2 content and the MDA content in leaves of creeping bentgrass in favor of water and redox homeostasis in summer. Based on the comprehensive evaluation of the subordinate function value analysis (SFVA), the CTS had the best effect on the mitigation of SBD, followed by GABA and NaCl in 2017, 2018, and 2019. The current study indicates that the foliar application of an appropriate dose of GABA, CTS, or NaCl provides a cost-effective strategy for mitigating SBD.

Elicitors directed in vitro growth and production of stevioside and other secondary metabolites in Stevia rebaudiana (Bertoni) Bertoni

Stevia rebaudiana (stevia) is a plant in the Asteraceae that contains several biologically active compounds including the antidiabetic diterpene glycosides (e.g. stevioside, rebaudioside and dulcoside) that can serve as zero-calorie sugar alternatives. In this study, an elicitation strategy was applied using 5% polyethylene glycol (PEG), sodium chloride (NaCl; 50 and 100 mM) and gibberellic acid (2.0 and 4.0 mg/L GA3) to investigate their effect on shoot morphogenesis, and the production of phenolics, flavonoids, total soluble sugars, proline and stevioside, as well as antioxidant activity, in shoot cultures of S. rebaudiana. Herewith, the media supplemented with 2 mg/L and 4 mg/L GA3 exhibited the highest shooting response (87% and 80%). The augmentation of lower concentrations of GA3 (2 mg/L) in combination with 6-benzylaminopurine (BAP) resulted in the maximum mean shoot length (11.1 cm). The addition of 100 mM NaCl salts to the media led to the highest observed total phenolics content (TPC; 4.11 mg/g-DW compared to the control 0.52 mg/g-DW), total flavonoids content (TFC; 1.26 mg/g-DW) and polyphenolics concentration (5.39 mg/g-DW) in shoots cultured. However, the maximum antioxidant activity (81.8%) was observed in shoots raised in media treated with 50 mM NaCl. The application of 2 mg/L of GA3 resulted in the highest accumulation of proline (0.99 μg/mL) as compared to controls (0.37 μg/mL). Maximum stevioside content (71 µL/mL) was observed in cultures supplemented with 100 mM NaCl and 5% PEG, followed by the 4 mg/L GA3 treatment (70 µL/mL) as compared to control (60 µL/mL). Positive correlation was observed between GA3 and stevioside content. Notably, these two compounds are derived from a shared biochemical pathway. These results suggest that elicitation is an effective option to enhance the accumulation of steviosides and other metabolites and provides the groundwork for future industrial scale production using bioreactors.