Search Results (19)

Plant growth is susceptible to abiotic stresses like salt and drought, and Na⁺/H⁺ antiporters (NHXs) play a pivotal role in stress responses. NHX proteins belong to the CPAs (cation/proton antiporters) family with a conserved Na⁺ (K⁺)/H⁺ exchange domain, which is widely involved in plant growth, development, and defense. While NHX genes have been extensively studied in model plants (e.g., Arabidopsis thaliana and Oryza sativa), research in other species remains limited. In this study, we identified nine NHX genes in foxtail millet (Setaria italica) and analyzed their systematic phylogeny, gene structure, protein characteristics, distribution of the chromosome, collinearity relationship, and cis-elements prediction at the promoter region. Phylogenetic analysis revealed that the members of the SiNHX gene family were divided into four subgroups. RT-qPCR analysis of the SiNHX family members showed that most genes were highly expressed in roots of foxtail millet, and their transcriptional levels responded to salt stress treatment. To determine SiNHX7’s function, we constructed overexpression Arabidopsis lines for each of the two transcripts of SiNHX7, and found that the overexpressed plants exhibited salt tolerance. These findings provide valuable insights for further study of the function of SiNHX genes and are of great significance for breeding new varieties of salt-resistant foxtail millet. Full article

Background:Citrus Exocortis Viroid (CEVd) is a non-coding RNA pathogen capable of infecting a wide range of plant species, despite its lack of protein-coding ability. Viroid infections induce significant alterations in various physiological and biochemical processes, particularly impacting plant metabolism. This study shows the metabolic changes upon viroid infection in tomato plants (Solanum lycopersicum var. ‘MoneyMaker’) exhibiting altered levels of salicylic acid (SA), a key signal molecule involved in the plant defence against this pathogen. Methods: Transgenic RNAi_S5H lines, which have the salicylic acid 5-hydroxylase gene silenced to promote SA accumulation, and NahG lines, which overexpress a salicylate hydroxylase to degrade SA into catechol and prevent its accumulation, were used to establish different SA levels in plants, resulting in varying degrees of resistance to viroid infection. The analysis was performed by using gas chromatography–mass spectrometry (GC-MS) to explore the role of volatile organic compounds (VOCs) in plant immunity against this pathogen. Results: Our results revealed distinct volatile profiles associated with plant immunity, where RNAi_S5H-resistant plants showed significantly enhanced production of monoterpenoids upon viroid infection. Moreover, viroid-susceptible NahG plants emitted a broad range of VOCs, whilst viroid-tolerant RNAi_S5H plants exhibited less variation in VOC emission. Conclusions: This study demonstrates that SA levels significantly influence metabolic responses and immunity in tomato plants infected by CEVd. The identification of differential emitted VOCs upon CEVd infection could allow the development of biomarkers for disease or strategies for disease control. Full article

Halotolerant plant growth-promoting bacteria (HT-PGPB) have attracted considerable attention for their significant potential in mitigating salt stress in crops. However, the current exploration and development of HT-PGPB remain insufficient to meet the increasing demands of agriculture. In this study, an HT-PGPB isolated from coastal saline-alkali soil in the Yellow River Delta was identified as Glutamicibacter endophyticus J2-5-19. The strain was capable of growing in media with up to 13% NaCl and producing proteases, siderophores, and the plant hormone IAA. Under 4‰ salt stress, inoculation with strain J2-5-19 significantly increased the wheat seed germination rate from 37.5% to 95%, enhanced the dry weight of maize seedlings by 41.92%, and notably improved the development of maize root systems. Moreover, this work presented the first whole-genome of Glutamicibacter endophyticus, revealing that G. endophyticus J2-5-19 resisted salt stress by expelling sodium ions and taking up potassium ions through Na⁺/H⁺ antiporters and potassium uptake proteins, while also accumulating compatible solutes such as betaine, proline, and trehalose. Additionally, the genome contained multiple key plant growth-promoting genes, including those involved in IAA biosynthesis, siderophore production, and GABA synthesis. The findings provide a theoretical foundation and microbial resources for the development of specialized microbial inoculants for saline-alkali soils. Full article

Submerged aquatic plants have potential applications in the phytoremediation of aquatic environments contaminated with arsenic (As). However, the role of epiphytic bacteria that grow on the surface of plants in As uptake and metabolism in plants has often been overlooked. An orthogonal experimental design with nine treatments, four factors, and three levels was conducted to inspect the effects of nitrogen (N, KNO₃, 2, 4, 10 mg/L), phosphorus (P, NaH₂PO₄·2H₂O, 0.02, 0.2, 1 mg/L), pH (6, 7, 9), and arsenate (As(V), Na₃AsO₄·12H₂O, 15, 75, 375 μg/L) on As accumulation and biotransformation in sterilized plants and to further explore the role of epiphytic bacteria in the metabolism of As by Hydrilla verticillata (L.f.) Royle. The results indicate that low N, intermediate P, and intermediate pH were beneficial for As accumulation (117.2 ± 62.2 μg/g DW) in sterilized plants, and epiphytic bacteria exhibited promotion (68%) in plants. High N promoted As absorption and transformation in non-sterilized plants but reduced As absorption in sterilized plants. Epiphytic bacteria in the medium showed significant As(III) oxidation, which was affected by environmental factors. These findings can promote remediation efficiency by regulating environmental factors for the phytoremediation of As-contaminated waters. Full article

Pepper (Capsicum annuum L.) fruit is a horticultural product consumed worldwide which has great nutritional and economic relevance. Besides the phenotypical changes that pepper fruit undergo during ripening, there are many associated modifications at transcriptomic, proteomic, biochemical, and metabolic levels. Nitric oxide (NO) is a recognized signal molecule that can exert regulatory functions in diverse plant processes including fruit ripening, but the relevance of NADPH as a fingerprinting of the crop physiology including ripening has also been proposed. Glucose-6-phosphate dehydrogenase (G6PDH) is the first and rate-limiting enzyme of the oxidative phase of the pentose phosphate pathway (oxiPPP) with the capacity to generate NADPH. Thus far, the available information on G6PDH and other NADPH-generating enzymatic systems in pepper plants, and their expression during the ripening of sweet pepper fruit, is very scarce. Therefore, an analysis at the transcriptomic, molecular and functional levels of the G6PDH system has been accomplished in this work for the first time. Based on a data-mining approach to the pepper genome and fruit transcriptome (RNA-seq), four G6PDH genes were identified in pepper plants and designated CaG6PDH1 to CaG6PDH4, with all of them also being expressed in fruits. While CaG6PDH1 encodes a cytosolic isozyme, the other genes code for plastid isozymes. The time-course expression analysis of these CaG6PDH genes during different fruit ripening stages, including green immature (G), breaking point (BP), and red ripe (R), showed that they were differentially modulated. Thus, while CaG6PDH2 and CaG6PDH4 were upregulated at ripening, CaG6PDH1 was downregulated, and CaG6PDH3 was slightly affected. Exogenous treatment of fruits with NO gas triggered the downregulation of CaG6PDH2, whereas the other genes were positively regulated. In-gel analysis using non-denaturing PAGE of a 50–75% ammonium-sulfate-enriched protein fraction from pepper fruits allowed for identifying two isozymes designated CaG6PDH I and CaG6PDH II, according to their electrophoretic mobility. In order to test the potential modulation of such pepper G6PDH isozymes, in vitro analyses of green pepper fruit samples in the presence of different compounds including NO donors (S-nitrosoglutathione and nitrosocysteine), peroxynitrite (ONOO⁻), a hydrogen sulfide (H₂S) donor (NaHS, sodium hydrosulfide), and reducing agents such as reduced glutathione (GSH) and L-cysteine (L-Cys) were assayed. While peroxynitrite and the reducing compounds provoked a partial inhibition of one or both isoenzymes, NaHS exerted 100% inhibition of the two CaG6PDHs. Taken together these data provide the first data on the modulation of CaG6PDHs at gene and activity levels which occur in pepper fruit during ripening and after NO post-harvest treatment. As a consequence, this phenomenon may influence the NADPH availability for the redox homeostasis of the fruit and balance its active nitro-oxidative metabolism throughout the ripening process. Full article

Anthropogenic climate change and species invasion are two major threats to biodiversity, affecting the survival and distribution of many species around the world. Studying the responses of invasive species under climate change can help better understand the ecological and genetic mechanisms of their invasion. However, the effects of warming and phosphorus deposition on the phenotype of native and invasive plants are unknown. To address the problem, we applied warming (+2.03 °C), phosphorus deposition (4 g m⁻² yr⁻¹ NaH₂PO₄), and warming × phosphorus deposition to Solidago canadensis and Artemisia argyi to measure the direct effects of environmental changes on growth and physiology at the seedling stage. Our results reveal that the physiology parameters of A. argyi and S. canadensis did not change significantly with the external environment. Under phosphorus deposition, S. canadensis had higher plant height, root length, and total biomass compared to A. argyi. Interestingly, warming has an inhibitory effect on the growth of both A. argyi and S. canadensis, but overall, the reduction in total biomass for S. canadensis (78%) is significantly higher than A. argyi (52%). When the two plants are treated with warming combined with phosphorus deposition, the advantage gained by S. canadensis from phosphorus deposition is offset by the negative effects of warming. Therefore, under elevated phosphorus, warming has a negative effect on the invasive S. canadensis and reduces its growth advantage. Full article

Although the effect of folic acid (FA) and its derivatives (folates) have been extensively studied in humans and animals, their effects are still unclear in most plant species, specifically under various abiotic stress conditions. Here, the impact of FA as a foliar application at 0, 0.1, and 0.2 mM was studied on snap bean seedlings grown under non-saline and salinity stress (50 mM NaCl) conditions. The results indicated that under salinity stress, FA-treated plants revealed a significant (p ≤ 0.05) increase in growth parameters (fresh and dry weight of shoot and root). A similar trend was observed in chlorophyll (Chl b), total chlorophyll, carotenoids, leaf relative water content (RWC), proline, free amino acids (FAA), soluble sugars, cell membrane stability index (CMSI), and K, Ca, and K/Na ratio compared to the untreated plants. In contrast, a significant decrease was observed in Na and salinity-induced oxidative damage as indicated by reduced H₂O₂ production (using biochemical and histochemical detection methods) and rate of lipid peroxidation (malondialdehyde; MDA). This enhancement was correlated by increasing the activities of antioxidant enzymes, i.e., superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (G-POX), and ascorbate peroxidase (APX). Gene expression analyses conducted using qRT-PCR demonstrated that genes coding for the Na⁺/H⁺ antiporter protein Salt Overly Sensitive 1 (SOS1), the tonoplast-localized Na⁺/H⁺ antiporter protein (NHX1), and the multifunctional osmotic protective protein (Osmotin) were significantly up-regulated in the FA-treated plants under both saline and non-saline treatments. Generally, treatment with 0.2 mM FA was more potent than 0.1 mM and can be recommended to improve snap bean tolerance to salinity stress. Full article

Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) is a cosmopolitan, highly polyphagous agricultural pest, which has the capacity to displace other native insect herbivores. Here, equipped with an integrated approach, the effect of developmental stages and feeding density of whiteflies on Myzus persicae performance in tobacco plants are investigated. Bioassay results showed that B. tabaci nymphs, but not adult, pre-infestation significantly reduced survival and fecundity of M. persicae, and the strongest resistance to M. persicae was detected at the medium density (9–10 nymphs/cm²). Neither low nor high feeding density of B. tabaci nymphs triggered visible resistance to aphids. However, no significant results were detected in salicylate-deficient NahG plants after B. tabaci nymph infestation. In addition to performance distinctions, hormone quantification and qPCR results revealed very different effects for nymph and adult whitefly stages on the defense responses in tobacco. B. tabaci nymph infestation significantly increased SA accumulation and SA-responsive genes (PR-1a, PR-2a) expression but suppressed JA-regulated responses. In contrast, tobacco plants responded to adult infestation by slightly increasing in both SA- and JA-regulated defenses. Furthermore, higher transcription level of Bt56, coding gene of a secretory salivary effector, was recorded in nymphs vs. adults, while silencing of Bt56 by virus-induced gene silencing (VIGS) partly impaired the aphid resistance induced by B. tabaci nymphs. These results proved that the induction of tobacco defense responses varied with the feeding stages of whiteflies: nymphs of B. tabaci, but not adults, induced a defense response against aphids, with a density threshold for this induced resistance. Full article

Recently, exogenous α-Lipoic acid (ALA) has been suggested to improve the tolerance of plants to a wide array of abiotic stresses. However, there is currently no definitive data on the role of ALA in wheat plants exposed to sodic alkaline stress. Therefore, this study was designed to evaluate the effects of foliar application by ALA at 0 (distilled water as control) and 20 µM on wheat seedlings grown under sodic alkaline stress (50 mM 1:1 NaHCO₃ & Na₂CO₃; pH 9.7. Under sodic alkaline stress, exogenous ALA significantly (p ≤ 0.05) improved growth (shoot fresh and dry weight), chlorophyll (Chl) a, b and Chl a + b, while Chl a/b ratio was not affected. Moreover, leaf relative water content (RWC), total soluble sugars, carotenoids, total soluble phenols, ascorbic acid, K and Ca were significantly increased in the ALA-treated plants compared to the ALA-untreated plants. This improvement was concomitant with reducing the rate of lipid peroxidation (malondialdehyde, MDA) and H₂O₂. Superoxide dismutase (SOD) and ascorbate peroxidase (APX) demonstrated greater activity in the ALA-treated plants compared to the non-treated ones. Conversely, proline, catalase (CAT), guaiacol peroxidase (G-POX), Na and Na/K ratio were significantly decreased in the ALA-treated plants. Under sodic alkaline stress, the relative expression of photosystem II (D2 protein; PsbD) was significantly up-regulated in the ALA treatment (67% increase over the ALA-untreated plants); while Δ pyrroline-5-carboxylate synthase (P5CS), plasma membrane Na⁺/H⁺ antiporter protein of salt overly sensitive gene (SOS1) and tonoplast-localized Na⁺/H⁺ antiporter protein (NHX1) were down-regulated by 21, 37 and 53%, respectively, lower than the ALA-untreated plants. These results reveal that ALA may be involved in several possible mechanisms of alkalinity tolerance in wheat plants. Full article

In plants, α-Lipoic acid (ALA) is considered a dithiol short-chain fatty acid with several strong antioxidative properties. To date, no data are conclusive regarding its effects as an exogenous application on salt stressed sorghum plants. In this study, we investigated the effect of 20 µM ALA as a foliar application on salt-stressed sorghum plants (0, 75 and 150 mM as NaCl). Under saline conditions, the applied-ALA significantly (p ≤ 0.05) stimulated plant growth, indicated by improving both fresh and dry shoot weights. A similar trend was observed in the photosynthetic pigments, including Chl a, Chl b and carotenoids. This improvement was associated with an obvious increase in the membrane stability index (MSI). At the same time, an obvious decrease in the salt induced oxidative damages was seen when the concentration of H₂O₂ and malondialdehyde (MDA) was reduced in the salt stressed leaf tissues. Generally, ALA-treated plants demonstrated higher antioxidant enzyme activity than in the ALA-untreated plants. A moderate level of salinity (75 mM) induced the highest activities of superoxide dismutase (SOD), guaiacol peroxidase (G-POX), and ascorbate peroxidase (APX). Meanwhile, the highest activity of catalase (CAT) was seen with 150 mM NaCl. Interestingly, applied-ALA led to a substantial decrease in the concentration of both Na and the Na/K ratio. In contrast, K and Ca exhibited a considerable increase in this respect. The role of ALA in the regulation of K⁺/Na⁺ selectivity under saline condition was confirmed through a molecular study (RT-PCR). It was found that ALA treatment downregulated the relative gene expression of plasma membrane (SOS1) and vacuolar (NHX1) Na⁺/H⁺ antiporters. In contrast, the high-affinity potassium transporter protein (HKT1) was upregulated. Full article

Plant protoplasts are significant for plant cell culture, somatic cell fusion, genetics, and breeding studies. In addition, in vitro plant regeneration has great importance for developmental biology, manifesting potential applications in agriculture and biotechnology. In this regard, we present a well-established protocol regarding protoplast isolation, cell culture and protoplast fusion of Jasminum spp. In particular, different tissues of Jasminum samab L. and Jasminum mesnyi were employed for protoplast isolation, and stem explants provided a high callus induction rate in a short period of time. The best source for protoplast isolation was calli tissues. The optimized isolation protocol consisted of digesting callus in an enzyme solution containing 0.4 M mannitol, 0.2 M MES, 1 M CaCl₂, 0.2 M KCL and 1 M NaH₂PO_4, 1.5% Cellulases onozuka R-10, 0.4% Macerozyme R-10 and 0.8% Pectinase for 4 h at 26 °C in the dark, providing a yield of 23.8 × 10⁶ Protoplast/gFW with 88% viability. Protoplasts were cultured both in liquid and agarose medium under optimum conditions, leading to microcalli formation after eight weeks. A 5% protoplast-fusion rate can be achieved when cultured in 40% (w/v) PEG-MW6000 supplemented with 0.1 M CaCl₂, 0.1 M sorbitol and 1 M Tris for 20 min. Furthermore, we developed an efficient PEG-mediated transformation protocol for jasmine protoplasts. The best results regarding protoplast transformation were obtained when the protoplast concentration was 4 × 10⁵ cells/mL and the exogenous plasmid DNA added had a concentration of 10 µg DNA/100 µL protoplast solution, followed by the application of 40% PEG-4000 for 10 min. Full article

Mineral nutrients are essential for plant growth and reproduction, yet only a few studies connect the nutritional status to plant innate immunity. The backbone of plant defense response is mainly controlled by two major hormones: salicylic acid (SA) and jasmonic acid (JA). This study investigated changes in the macronutrient concentration (deficiency/excess of nitrogen, phosphorus, potassium, magnesium, and sulfur) on the expression of PR1, a well-characterized marker in the SA-pathway, and PDF1.2 and LOX2 for the JA-pathway, analyzing plants carrying the promoter of each gene fused to GUS as a reporter. After histochemical GUS assays, we determined that PR1 gene was strongly activated in response to sulfur (S) deficiency. Using RT-PCR, we observed that the induction of PR1 depended on the function of Non-expressor of Pathogenesis-Related gene 1 (NPR1) and SA accumulation, as PR1 was not expressed in npr1-1 mutant and NahG plants under S-deprived conditions. Plants treated with different S-concentrations showed that total S-deprivation was required to induce SA-mediated defense responses. Additionally, bioassays revealed that S-deprived plants, induced resistance to the hemibiotrophic pathogen Pseudomonas syringae pv. DC3000 and increase susceptibility to the necrotrophic Botrytis cinerea. In conclusion, we observed a relationship between S and SA/JA-dependent defense mechanisms in Arabidopsis. Full article

Salicylic acid (SA) is well known hormonal molecule involved in cell death regulation. In response to a broad range of environmental factors (e.g., high light, UV, pathogens attack), plants accumulate SA, which participates in cell death induction and spread in some foliar cells. LESION SIMULATING DISEASE 1 (LSD1) is one of the best-known cell death regulators in Arabidopsis thaliana. The lsd1 mutant, lacking functional LSD1 protein, accumulates SA and is conditionally susceptible to many biotic and abiotic stresses. In order to get more insight into the role of LSD1-dependent regulation of SA accumulation during cell death, we crossed the lsd1 with the sid2 mutant, caring mutation in ISOCHORISMATE SYNTHASE 1(ICS1) gene and having deregulated SA synthesis, and with plants expressing the bacterial nahG gene and thus decomposing SA to catechol. In response to UV A+B irradiation, the lsd1 mutant exhibited clear cell death phenotype, which was reversed in lsd1/sid2 and lsd1/NahG plants. The expression of PR-genes and the H₂O₂ content in UV-treated lsd1 were significantly higher when compared with the wild type. In contrast, lsd1/sid2 and lsd1/NahG plants demonstrated comparability with the wild-type level of PR-genes expression and H₂O₂. Our results demonstrate that SA accumulation is crucial for triggering cell death in lsd1, while the reduction of excessive SA accumulation may lead to a greater tolerance toward abiotic stress. Full article

Cotton is an important economic crop affected by different abiotic stresses at different developmental stages. Salinity limits the growth and productivity of crops worldwide. Na⁺/H⁺ antiporters play a key role during the plant development and in its tolerance to salt stress. The aim of the present study was a genome-wide characterization and expression pattern analysis under the salinity stress of the sodium-proton antiporter (NHX) of Gossypium barbadense in comparison with Gossypium hirsutum. In G. barbadense, 25 NHX genes were identified on the basis of the Na⁺_H⁺ exchanger domain. All except one of the G. barbadense NHX transporters have an Amiloride motif that is a known inhibitor of Na⁺ ions in plants. A phylogenetic analysis inferred three classes of GbNHX genes—viz., Vac (GbNHX1, 2 and 4), Endo (GbNHX6), and PM (GbNHX7). A high number of the stress-related cis-acting elements observed in promoters show their role in tolerance against abiotic stresses. The Ka/Ks values show that the majority of GbNHX genes are subjected to strong purifying selection under the course of evolution. To study the functional divergence of G. barbadense NHX transporters, the real-time gene expression was analyzed under salt stress in the root, stem, and leaf tissues. In G. barbadense, the expression was higher in the stem, while in G. hirsutum the leaf and root showed a high expression. Moreover, our results revealed that NHX2 homologues in both species have a high expression under salinity stress at higher time intervals, followed by NHX7. The protein-protein prediction study revealed that GbNHX7 is involved in the CBL-CIPK protein interaction pathway. Our study also provided valuable information explaining the molecular mechanism of Na⁺ transport for the further functional study of Gossypium NHX genes. Full article

Fatty acids (FAs) have been implicated in signaling roles in plant defense responses. We previously reported that mutation or RNAi-knockdown (OsSSI2-kd) of the rice OsSSI2 gene, encoding a stearoyl acyl carrier protein FA desaturase (SACPD), remarkably enhanced resistance to blast fungus Magnaporthe oryzae and the leaf-blight bacterium Xanthomonas oryzae pv. oryzae (Xoo). Transcriptomic analysis identified six AAA-ATPase family genes (hereafter OsAAA-ATPase1–6) upregulated in the OsSSI2-kd plants, in addition to other well-known defense-related genes. Here, we report the functional analysis of OsAAA-ATPase1 in rice’s defense response to M. oryzae. Recombinant OsAAA-ATPase1 synthesized in Escherichia coli showed ATPase activity. OsAAA-ATPase1 transcription was induced by exogenous treatment with a functional analogue of salicylic acid (SA), benzothiadiazole (BTH), but not by other plant hormones tested. The transcription of OsAAA-ATPase1 was also highly induced in response to M. oryzae infection in an SA-dependent manner, as gene induction was significantly attenuated in a transgenic rice line expressing a bacterial gene (nahG) encoding salicylate hydroxylase. Overexpression of OsAAA-ATPase1 significantly enhanced pathogenesis-related gene expression and the resistance to M. oryzae; conversely, RNAi-mediated suppression of this gene compromised this resistance. These results suggest that OsAAA-APTase1 plays an important role in SA-mediated defense responses against blast fungus M. oryzae. Full article