Biological Activity and Phytochemical Characteristics of Star Anise (Illicium verum) Essential Oil and Its Anti-Salmonella Activity on Sous Vide Pumpkin Model.

Illicium verum, commonly known as star anise, represents one of the notable botanical species and is recognized for its rich reservoir of diverse bioactive compounds. Beyond its culinary application as a spice, this plant has been extensively utilized in traditional medicine. Given the contemporary emphasis on incorporating natural resources into food production, particularly essential oils, to enhance sensory attributes and extend shelf life, our study seeks to elucidate the chemical composition and evaluate the antibacterial (in vitro, in situ) and insecticidal properties of Illicium verum essential oil (IVEO). Also, microbiological analyses of pumpkin sous vide treated with IVEO after inoculation of Salmonella enterica were evaluated after 1 and 7 days of study. GC/MS analysis revealed a significantly high amount of (E)-anethole (88.4%) in the investigated EO. The disc diffusion method shows that the antibacterial activity of the IVEO ranged from 5.33 (Streptococcus constellatus) to 10.33 mm (Citrobacter freundii). The lowest minimal inhibition concentration was found against E. coli and the minimum biofilm inhibition concertation was found against S. enterica. In the vapor phase, the best antimicrobial activity was found against E. coli in the pears model and against S. sonei in the beetroot model. The application of the sous vide method in combination with IVEO application decreased the number of microbial counts and eliminated the growth of S. enterica. The most isolated microbiota identified from the sous vide pumpkin were Bacillus amyloliquefaciens, B. cereus, B. licheniformis, and Ralstonia picketii. Modifications to the protein composition of biofilm-forming bacteria S. enterica were suggested by the MALDI TOF MS instigations. The IVEO showed insecticidal potential against Harmonia axyridis. Thanks to the properties of IVEO, our results suggest it can be used in the food industry as a natural supplement to extend the shelf life of foods and as a natural insecticide.

Citrus limon Essential Oil: Chemical Composition and Selected Biological Properties Focusing on the Antimicrobial (In Vitro, In Situ), Antibiofilm, Insecticidal Activity and Preservative Effect against Salmonella enterica Inoculated in Carrot.

New goals for industry and science have led to increased awareness of food safety and healthier living in the modern era. Here, one of the challenges in food quality assurance is the presence of pathogenic microorganisms. As planktonic cells can form biofilms and go into a sessile state, microorganisms are now more resistant to broad-spectrum antibiotics. Due to their proven antibacterial properties, essential oils represent a potential option to prevent food spoilage in the search for effective natural preservatives. In this study, the chemical profile of Citrus limon essential oil (CLEO) was evaluated. GC-MS analysis revealed that limonene (60.7%), ß-pinene (12.6%), and γ-terpinene (10.3%) are common constituents of CLEO, which prompted further research on antibacterial and antibiofilm properties. Minimum inhibitory concentration (MIC) values showed that CLEO generally exhibits acceptable antibacterial properties. In addition, in situ antimicrobial research revealed that vapour-phase CLEO can arrest the growth of Candida and Y. enterocolitica species on specific food models, indicating the potential of CLEO as a preservative. The antibiofilm properties of CLEO were evaluated by MIC assays, crystal violet assays, and MALDI-TOF MS analysis against S. enterica biofilm. The results of the MIC and crystal violet assays showed that CLEO has strong antibiofilm activity. In addition, the data obtained by MALDI-TOF MS investigation showed that CLEO altered the protein profiles of the bacteria studied on glass and stainless-steel surfaces. Our study also found a positive antimicrobial effect of CLEO against S. enterica. The anti-Salmonella activity of CLEO in vacuum-packed sous vide carrot samples was slightly stronger than in controls. These results highlight the advantages of the antibacterial and antibiofilm properties of CLEO, suggesting potential applications in food preservation.

Mixture design of α-pinene, α-terpineol, and 1,8-cineole: A multiobjective response followed by chemometric approaches to optimize the antibacterial effect against various bacteria and antioxidant activity.

α-Pinene, α-terpineol, and 1,8-cineole are compounds naturally present in essential oils, although their amounts vary from oil to oil. Although several studies have reported their antibacterial and antioxidant effects, there are few reports on the synergistic or antagonistic effects of their combinations. The objective of this study was to investigate the combined antibacterial effect of these three compounds. To our knowledge, this is the first report on the prediction of their optimal combination using the mixture design approach. The experimental antibacterial activity of the α-pinene, α-terpineol, and 1,8-cineole mixtures depended on the proportion of each compound in the mixture and the target strain, with minimum inhibitory concentrations (MIC) ranging from 0.31 to 1.85 mg/mL. Using the increased simplex-centroid mixture design, the mixture containing 0.33% of each molecule proved to be the most effective against Bacillus cereus and had the lowest MIC values. In addition, α-pinene, α-terpineol, and 1,8-cineole showed significant antioxidant activity against 2,2-picryl-1-hydrazyl radical (DPPH), with IC50 values of 24.53 ± 0.05, 65.63 ± 0.71, and 63.58 ± 0.01 µg/mL, respectively. Statistical planning and the development of utility profiles of the substance mixtures can predict the optimal composition that will exhibit the highest antibacterial activity against B. cereus as well as antioxidant properties. Furthermore, the synergistic effect of the mixtures can contribute significantly to their successful use as natural preservatives in various applications.

GASA Proteins: Review of Their Functions in Plant Environmental Stress Tolerance.

Gibberellic acid-stimulated Arabidopsis (GASA) gene family is a class of functional cysteine-rich proteins characterized by an N-terminal signal peptide and a C-terminal-conserved GASA domain with 12 invariant cysteine (Cys) residues. GASA proteins are widely distributed among plant species, and the majority of them are involved in the signal transmission of plant hormones, the regulation of plant development and growth, and the responses to different environmental constraints. To date, their action mechanisms are not completely elucidated. This review reports an overview of the diversity, structure, and subcellular localization of GASA proteins, their involvement in hormone crosstalk and redox regulation during development, and plant responses to abiotic and biotic stresses. Knowledge of this complex regulation can be a contribution to promoting multiple abiotic stress tolerance with potential agricultural applications through the engineering of genes encoding GASA proteins and the production of transgenic plants.

Recent Advances in the Incorporation of Polysaccharides with Antioxidant and Antibacterial Functions to Preserve the Quality and Shelf Life of Meat Products.

Meat and meat products are susceptible to various types of natural processes such as oxidative degradation due to their high content of protein and essential amino acids. However, finding solutions to maintain the nutritional and sensory quality of meat and meat products is unavoidable. Hence, there is a pressing need to investigate alternatives to synthetic preservatives, focusing on active biomolecules of natural provenance. Polysaccharides are natural polymers of various sources that exhibit antibacterial and antioxidant properties via a variety of mechanisms, owing to their diversity and structural variation. For this reason, these biomolecules are widely studied in order to improve texture, inhibit the growth of pathogens, and improve the oxidative stability and sensory characteristics of meat products. However, the literature has not addressed their biological activity in meat and meat products. This review summarizes the various sources of polysaccharides, their antioxidant and antibacterial activities (mainly against pathogenic food strains), and their use as natural preservatives to replace synthetic additives in meat and meat products. Special attention is given to the use of polysaccharides to improve the nutritional value of meat, resulting in more nutrient-rich meat products with higher polysaccharide content and less salt, nitrites/nitrates, and cholesterol.

Functional Characterization of Lobularia maritima LmTrxh2 Gene Involved in Cold Tolerance in Tobacco through Alleviation of ROS Damage to the Plasma Membrane.

Cold stress is a key environmental factor affecting plant growth and development, crop productivity, and geographic distribution. Thioredoxins (Trxs) are small proteins that are ubiquitously expressed in all organisms and implicated in several cellular processes, including redox reactions. However, their role in the regulation of cold stress in the halophyte plant Lobularia maritima remains unknown. We recently showed that overexpression of LmTrxh2, which is the gene that encodes the h-type Trx protein previously isolated from L. maritima, led to an enhanced tolerance to salt and osmotic stress in transgenic tobacco. This study functionally characterized the LmTrxh2 gene via its overexpression in tobacco and explored its cold tolerance mechanisms. Results of the RT-qPCR and western blot analyses indicated differential temporal and spatial regulation of LmTrxh2 in L. maritima under cold stress at 4 °C. LmTrxh2 overexpression enhanced the cold tolerance of transgenic tobacco, as evidenced by increased germination rate, fresh weight and catalase (CAT), superoxide dismutase (SOD) and peroxidase (POD) activities; reduced malondialdehyde levels, membrane leakage, superoxide anion (O2-), and hydrogen peroxide (H2O2) levels; and higher retention of chlorophyll than in non-transgenic plants (NT). Furthermore, the transcript levels of reactive oxygen species (ROS)-related genes (NtSOD and NtCAT1), stress-responsive late embryogenis abundant protein 5 (NtLEA5), early response to dehydration 10C (NtERD10C), DRE-binding proteins 1A (NtDREB1A), and cold-responsive (COR) genes (NtCOR15A, NtCOR47, and NtKIN1) were upregulated in transgenic lines compared with those in NT plants under cold stress, indicating that LmTrxh2 conferred cold stress tolerance by enhancing the ROS scavenging ability of plants, thus enabling them to maintain membrane integrity. These results suggest that LmTrxh2 promotes cold tolerance in tobacco and provide new insight into the improvement of cold-stress resistance to cold stress in non-halophyte plants and crops.

The Essential Oil of Tunisian Halophyte Lobularia maritima: A Natural Food Preservative Agent of Ground Beef Meat.

This study was directed towards the investigation of the chemical composition and antimicrobial properties of the essential oil of Tunisian halophyte Lobularia maritime (LmEO). The antibacterial effects against major food-borne pathogenic and food spoilage bacteria were tested using the well diffusion method, followed by the determination of the minimum inhibitory (MIC) and bactericidal (MBC) concentrations. The essential oil has shown strong antimicrobial activity against eight pathogenic strains, which was attributed mostly to predominant constituents of the essential oil: benzyl alcohol, linalool, terpien-4-ol and globulol, as well as to synergistic effects of its major and minor constituents. Considering strong antimicrobial effects of the tested essential oil, it was further tested as a natural alternative to food preservatives, using minced beef meat as a model system. Minced beef meat was spiked with 0.019, 0.038, and 0.076% of the essential oil and stored during 14 days at 4 °C, monitoring its microbiological, physicochemical, and sensory properties. Chemical analyses revealed that meat treated with 0.076% of LmEO at underwent a significant decrease (p < 0.05) in primary and secondary lipid oxidation and reduced metmyoglobin accumulation compared with control samples. Furthermore, microflora proliferation in the meat model system spiked with 0.076% of LmEO was significantly (p < 0.05) reduced in comparison to control. In addition, two multivariate exploratory techniques, namely principal component analysis (PCA) and hierarchical analysis (HCA), were applied to the obtained data sets to describe the relationship between the main characteristics of the meat samples with and without essential oil addition. The chemometric approach highlighted the relationships between meat quality parameters. Overall, results indicated that the essential oil of Lobularia maritima deserves to be considered as a natural preservative in the meat industry.

Antioxidant and hepatoprotective effects of novel heteropolysaccharide isolated from Lobularia maritima on CCl4-induced liver injury in rats.

The aim of the present study was to investigate the extraction and the characterization of a novel heteropolysaccharide from Tunisian halophyte Lobularia maritima (LmPS). We were also interested in its antioxidant, anti-inflammatory, and hepatoprotective effects on carbon tetrachloride (CCl4)-induced liver injury in rats. LmPS physicochemical properties were evaluated by thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), thermogravimetric analysis (TGA), and UV absorption. According to TLC and HPLC results, LmPS was a heteropolysaccharide composed of glucose, galactose, and xylose. Its molecular weight was 130.62 kDa. This heteropolysaccharide was characterized by a significant antioxidant potential and was efficient against oxidative stress and CCL4-induced hepatotoxicity in rat Wistar models (n = 8) treated with a single dose of LmPS 250 mg/kg of body weight. This was evidenced by a significant increase in serum marker enzymes specially aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH). The cytokines released after stimulation of rats with LmPS showed high anti-inflammatory profiles with an increased rate of interleukine-10 (IL-10) with 0.03 pg/mL compared to animals treated only with CCl4. On the contrary, we noticed a decrease of the other cytokines (tumor necrosis factor α: TNF-α, interleukine-6: IL-6, transforming growth factor beta 1: TGF-ß1) with average concentration values of <0.2, 0.1, and 0.04 pg/mL, respectively. Besides, histopathological examinations revealed that CCl4 causes acute liver damage, characterized by extensive hepatocellular necrosis, vacuolization, and inflammatory cell infiltration, as well as DNA fragmentation. LmPS administration at a dose of 250 mg/kg resulted in a significant hepatoprotection, evidenced by a reduction of CCl4-induced oxidative damage for all tested markers. These findings eagerly confirmed that LmPS was effective in the protection against CCl4-induced hepatotoxicity and genotoxicity. It, therefore, suggested a potential therapeutic use of this polysaccharide as an alternative medicine for patients with acute liver diseases.

Lobularia maritima leave extract, a nutraceutical agent with antioxidant activity, protects against CCl4-induced liver injury in mice.

Lobularia maritima (Alyssum maritimum, Brassicaceae), commonly known as sweet alyssum, is an annual ornamental halophyte widely spread along the Tunisian seashore. Lobularia maritima leaf ethanol extract was tested in an experimental model of hepatotoxicity induced by carbon tetrachloride (CCl4). L. maritima extract was found to possess in vitro antioxidant activity by scavenging the DPPH radical (IC50= 45 µg/mL), reducing/chelating iron ions and inhibiting liver lipid peroxidation induced by FeSO4. The levels of total phenolics and flavonoids were 175 ± 2.66 mg GAE/g, and 35 ± 2.88 mg QE/g respectively. Moreover, HPLC analysis revealed six compounds, namely gallic, salicylic, ellagic and ferulic acids as well as catechin and quercetin. A mice model of acute liver injury was successfully established after a single intraperitoneal injection of CCl4, as evidenced by histological analysis, Masson trichrome and Sirius red staining. Compared with the CCl4 intoxicated group, the L. maritima treatment resulted to reduce the liver serum marker enzymes, lipid peroxidation and increased the activities of antioxidant enzymes with further amelioration in the oxidative stress. The present findings discover the therapeutic potentials of L. maritima empowered with promising natural leads for the treatment of oxidative stress associated health disorders by attenuating free radicals, inhibiting lipid peroxidation, and upregulating the tissue-specific antioxidant enzymes.

Novel non-specific lipid-transfer protein (TdLTP4) isolated from durum wheat: Antimicrobial activities and anti-inflammatory properties in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages.

Lipid transfer proteins (LTP) are members of the family of pathogenesis-related proteins (PR-14) that play a key role in plant defense mechanisms. In this study, a novel gene TdLTP4 encoding an antifungal protein from wheat (cv. Om Rabiaa) was cloned, overexpressed in Escherichia coli BL-21 (DE3) and enriched using ammonium sulfate fractionation. The TdLTP4 fusion protein was then tested against a panel of pathogens, food-borne and spoilage bacteria and fungi in order to evaluate the antimicrobial properties. TdLTP4 was applied to 0.5 µg/mL LPS-induced RAW 264.7 macrophages in vitro at different concentrations (5, 10, 20, 50 and 100 µg/mL). Levels of nitric oxide (NO), pro-inflammatory cytokines interleukin (IL)-1ß (IL-1 ß), interleukin (IL)-6 (IL-6), tumor necrosis factor (TNF-α) and anti-inflammatory cytokine IL-10 in the supernatant fraction were measured using enzyme-linked immunosorbent assay (ELISA). Expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) were detected via Western blot. The inhibition zones and minimal inhibitory concentration (MIC) values of bacterial strains were in the range of 14-26 mm and 62.5-250 µg/mL, respectively. Moreover, a remarkable activity against several fungal strains was revealed. TdLTP4 (5-100 µg/mL) decreased the production of NO (IC50 = 4.32 µg/mL), IL-6 (IC50 = 11.52 µg/mL), IL-1ß (IC50 = 7.87 µg/mL) and TNF-α (IC50 = 8.66 µg/mL) by lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. TdLTP4 could modulate the macrophages inflammatory mode by causing reduction in iNOS and COX-2. According to these findings, TdLTP4 fusion protein could be used as natural anti-inflammatory and antimicrobial agent in food preservation and human health.

Characterization of a novel LmSAP gene promoter from Lobularia maritima: Tissue specificity and environmental stress responsiveness.

Halophyte Lobularia maritima LmSAP encodes an A20AN1 zinc-finger stress-associated protein which expression is up-regulated by abiotic stresses and heavy metals in transgenic tobacco. To deepen our understanding of LmSAP function, we isolated a 1,147 bp genomic fragment upstream of LmSAP coding sequence designated as PrLmSAP. In silico analyses of PrLmSAP revealed the presence of consensus CAAT and TATA boxes and cis-regulatory elements required for abiotic stress, phytohormones, pathogen, and wound responses, and also for tissue-specific expression. The PrLmSAP sequence was fused to the ß-glucuronidase (gusA) reporter gene and transferred to rice. Histochemical GUS staining showed a pattern of tissue-specific expression in transgenic rice, with staining observed in roots, coleoptiles, leaves, stems and floral organs but not in seeds or in the root elongation zone. Wounding strongly stimulated GUS accumulation in leaves and stems. Interestingly, we observed a high stimulation of the promoter activity when rice seedlings were exposed to NaCl, PEG, ABA, MeJA, GA, cold, and heavy metals (Al3+, Cd2+, Cu2+ and Zn2+). These results suggest that the LmSAP promoter can be a convenient tool for stress-inducible gene expression and is a potential candidate for crop genetic engineering.

Early effects of salt stress on the physiological and oxidative status of the halophyte Lobularia maritima.

Soil salinity is an abiotic stress that reduces agricultural productivity. For decades, halophytes have been studied to elucidate the physiological and biochemical processes involved in alleviating cellular ionic imbalance and conferring salt tolerance. Recently, several interesting genes with proven influence on salt tolerance were isolated from the Mediterranean halophyte Lobularia maritima (L.) Desv. A better understanding of salt response in this species is needed to exploit its potential as a source of stress-related genes. We report the characterisation of L. maritima's response to increasing NaCl concentrations (100-400 mM) at the physiological, biochemical and molecular levels. L. maritima growth was unaffected by salinity up to 100 mM NaCl and it was able to survive at 400 mM NaCl without exhibiting visual symptoms of damage. Lobularia maritima showed a Na+ and K+ accumulation pattern typical of a salt-includer halophyte, with higher contents of Na+ in the leaves and K+ in the roots of salt-treated plants. The expression profiles of NHX1, SOS1, HKT1, KT1 and VHA-E1 in salt-treated plants matched this Na+ and K+ accumulation pattern, suggesting an important role for these transporters in the regulation of ion homeostasis in leaves and roots of L. maritima. A concomitant stimulation in phenolic biosynthesis and antioxidant enzyme activity was observed under moderate salinity, suggesting a potential link between the production of polyphenolic antioxidants and protection against salt stress in L. maritima. Our findings indicate that the halophyte L. maritima can rapidly develop physiological and antioxidant mechanisms to adapt to salt and manage oxidative stress.

A Lobularia maritima LmSAP protein modulates gibberellic acid homeostasis via its A20 domain under abiotic stress conditions.

Stress-associated proteins (SAPs) are favorable targets to improve stress tolerance in plants, owing to their roles in developmental processes and stress responses. However, the role of SAPs and the molecular mechanisms by which they regulate plant stress responses remain poorly understood. Previously, it was reported that LmSAP expression was upregulated by various abiotic stressors in Lobularia maritima, and that transgenic tobacco lines with constitutively expressed LmSAPΔA20 and LmSAPΔA20-ΔAN1 showed dwarf phenotypes due to the deficiency of cell elongation under salt and osmotic stresses. In this study, we examined the function of A20 domain in the GA pathway in response to abiotic stresses. Transient expression of acGFP-LmSAPΔA20 and acGFP-LmSAPΔA20-ΔAN1 in onion epidermal cells demonstrated that these fused proteins were localized in the nucleo-cytoplasm. However, the truncated form acGFP-LmSAPΔAN1 was localized in the nucleus. Moreover, comparison of native and truncated LmSAP showed dramatic structural changes caused by the deletion of the A20 domain, leading to loss of function and localization. Interestingly, overexpression LmSAP and truncated LmSAPΔAN1 led to up-regulation of GA biosynthetic genes and increased total gibberellins (GAs) content, corresponding with accelerated development in transgenic tobacco plants. Moreover, the dwarf phenotype of the transgenic lines that express LmSAPΔA20 and LmSAPΔA20-ΔAN1 under stress conditions was fully restored by the application of exogenous GA3. These findings improve our understanding of the role of LmSAP in regulating GA homeostasis, which is important for regulating plant development under abiotic stress conditions.

A novel Triticum durum Annexin 12 protein: Expression, purification and biological activities against Listeria monocytogenes growth in meat under refrigeration.

The present study was undertaken to investigate the expression, purification and biological activities of a novel Triticum durum Annexin 12 protein (TdAnn12). The findings indicated that the molecular weight of the purified TdAnn12 was estimated to 35 kDa. The purified TdAnn12 protein was modulated by, Methyl-jasmonate, and ethephon treatments. The purified TdAnn12 protein displayed good antimicrobial activities against 9 tested pathogenic bacteria. The antioxidant activities showed that TdAnn12 displayed an excellent DPPH scavenging ability with an IC50 of 8.33 µg/ml and a strong Beta-carotene bleaching inhibition after 120 min of incubation with an IC50 of 2 µg/ml the cytotoxic effects of the TdAnn 12 showed that HepG2 and MCF-7 were examined by MTT assay. The IC50 values were 250.35 and 400.25 µg/ml for HepG2 and MCF-7 cells, respectively. The inhibitory effects of this TdAnn12 was assessed in vivo against Listeria monocytogenes, inoculated in minced beef meat at 6.105 CFU/g amended with different concentrations of the purified TdAnn12 and stored at 4 °C for 21 days. Results showed an excellent inhibitory effect of TdAnn12 of this pathogenic bacterium at 4 °C. Overall, the TdAnn12 have potential application as active ingredients in food and pharmaceutical industry.

Overexpression of Triticum durum TdAnn12 gene confers stress tolerance through scavenging reactive oxygen species in transgenic tobacco.

Plant annexins are proteins with multiple functions and roles in plant development and responses to abiotic stresses. We report here the functional analysis of the TdAnn12 annexin protein isolated from Triticum durum Desf. We have previously shown that TdAnn12 expression is highly induced by different abiotic stresses. In the present study, to investigate the physiological and biochemical stress-induced responses, we overexpressed TdAnn12 in tobacco. We demonstrate that transgenic tobacco plants expressing TdAnn12 exhibited enhanced tolerance to salt, osmotic stress and H2O2 at the seedling stage. Under greenhouse conditions, these plants showed tolerance to drought and salt stresses. Moreover, scavenging reactive oxygen species (ROS), higher chlorophyll content, lower lipid peroxidation levels and increased antioxidant activities (peroxidase, catalase and superoxide dismutase) were observed. Finally, accumulation of TdAnn12 in tobacco positively affects the regulation of some stress-related genes (MnSOD, APX1, CAT1, P5CS, NHX1, SOS1 and DREB1A). TdAnn12 interacts directly or indirectly with stress-related genes that could stimulate an adaptive potential to gain tolerance which is not present in non-transgenic (NT) plants. Our results clearly show that overexpression of TdAnn12 in transgenic tobacco improves stress tolerance through the removal of ROS.

Functional domain analysis of LmSAP protein reveals the crucial role of the zinc-finger A20 domain in abiotic stress tolerance.

Stress-associated proteins (SAPs), such as A20/AN1 zinc-finger domain-containing proteins, have emerged as a novel class of proteins involved in abiotic stress signaling, and they are important candidates for preventing the loss of yield caused by exposure to environmental stresses. In a previous report, it was found that the ectopic-expression of Lobularia maritima stress-associated protein, LmSAP, conferred tolerance to abiotic and heavy metal stresses in transgenic tobacco plants. This study aimed to investigate the functions of the A20 and AN1 domains of LmSAP in salt and osmotic stress tolerance. To this end, in addition to the full-length LmSAP gene, we have generated three LmSAP-truncated forms (LmSAPΔA20, LmSAPΔAN1, and LmSAPΔA20-ΔAN1). Heterologous expression in Saccharomyces cerevisiae of different truncated forms of LmSAP revealed that the A20 domain is essential to increase cell tolerance to salt, ionic, and osmotic stresses. Transgenic tobacco plants overexpressing LmSAP and LmSAPΔAN1 constructs exhibited higher tolerance to salt and osmotic stresses in comparison to the non-transgenic plants (NT) and lines transformed with LmSAPΔA20 and LmSAPΔA20-ΔAN1 constructs. Similarly, transgenic plants overexpressing the full-length LmSAP gene and LmSAPΔAN1 truncated domain maintained higher superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) enzymatic activities due to the high expression levels of the genes encoding these key antioxidant enzymes, MnSOD, POD, and CAT1, as well as accumulated lower levels of malondialdehyde (MDA) under salt and osmotic stresses compared to NT and LmSAPΔA20 and LmSAPΔA20-ΔAN1 forms. These findings provide insights into the pivotal role of A20 and AN1 domains of LmSAP protein in salt and osmotic stress tolerance.

Stress associated protein from Lobularia maritima: Heterologous expression, antioxidant and antimicrobial activities with its preservative effect against Listeria monocytogenes inoculated in beef meat.

Stress associated Protein from Lobularia maritima (LmSAP) transcripts showed up-regulation in response to wounding, Methyl-jasmonate acid, Picolinic acid, and ethephon treatments. The antioxidant activities of the purified LmSAP protein were evaluated using various in vitro and in vivo assays. LmSAP has an excellent DPPH scavenging ability and exhibits notable reducing power capacity and led to a decrease in TBARS values. The LmSAP showed an important antimicrobial activity against different pathogenic strains tested. In fact, the Inhibition Zone (IZ), Minimal Inhibitory Concentration (MIC) and The Minimum Bactericidal Concentration (MCB) recorded were in the range of 14-22 mm, 62-250 µg/ml and 125-250 µg/ml, respectively. The LmSAP protein successfully inhibited the growth of L. monocytogenes in minced beef meat. Furthermore, the LmSAP enhanced the sensory acceptability extending the shelf life of minced beef meat. Overall, the finding demonstrates that the application of LmSAP may open new promising opportunities for the prevention of bacterial contamination, particularly L. monocytogenes, during minced beef meat storage.

Potential anti-inflammatory and antioxidant effects of Citrus aurantium essential oil against carbon tetrachloride-mediated hepatotoxicity: A biochemical, molecular and histopathological changes in adult rats.

The present study aimed (1) to investigate the chemical composition as well as the anti-inflammatory properties and in vitro antioxidant activity of Citrus aurantium peel essential oil (pEOCa) and (2) to evaluate its potential effect in vivo. The main results showed that the major components of pEOCa are Limonene and Linalool. Additionally, DPPH scavenging ability and ß-carotene bleaching inhibition tests confirmed the antioxidant capacity of pEOCa. Our oil reduced the production of NO by LPS-stimulated RAW264,7 macrophages in a concentration-dependent. This inhibition occurred at a transcriptional level. pEOCa in CCl4 treated rats alleviated hepatotoxicity as monitored by the improvement of hepatic oxidative stress biomarkers levels plasma biochemical parameters, and DNA molecule aspect. Furthermore, the mRNA gene expression of Cu-Zn SOD, CAT, and GPx increased under CCl4 + pEOCa exposure to reach the same value to the control. Similarly, antioxidant activities of these three enzymes changed in accordance with the mRNA levels. These results were confirmed by the histological results. It seems obvious that the treatment with pEOCa prevented liver damage induced by CCl4 , thus preventing the harmful effects of free radicals.

Overexpression of AlTMP2 gene from the halophyte grass Aeluropus littoralis in transgenic tobacco enhances tolerance to different abiotic stresses by improving membrane stability and deregulating some stress-related genes.

Herein, we report isolation of the AlTMP2 gene from the halophytic C4 grass Aeluropus littoralis. The subcellular localization suggested that AlTMP2 is a plasma membrane protein. In A. littoralis exposed to salt and osmotic stresses, the AlTMP2 gene was induced early and at a high rate, but was upregulated relatively later in response to abscisic acid and cold treatments. Expression of AlTMP2 in tobacco conferred improved tolerance against salinity, osmotic, H2O2, heat, and freezing stresses at the germination and seedling stages. Under control conditions, no growth or yield penalty were mentioned in transgenic plants due to the constitutive expression of AlTMP2. Interestingly, under greenhouse conditions, the seed yield of transgenic plants was significantly higher than that of non-transgenic (NT) plants grown under salt or drought stress. Furthermore, AlTMP2 plants had less electrolyte leakage, higher membrane stability, and lower Na+ and higher K+ accumulation than NT plants. Finally, six stress-related genes were shown to be deregulated in AlTMP2 plants relative to NT plants under both control and stress conditions. Collectively, these results indicate that AlTMP2 confers abiotic stress tolerance by improving ion homeostasis and membrane integrity, and by deregulating certain stress-related genes.

The LmSAP gene isolated from the halotolerant Lobularia maritima improves salt and ionic tolerance in transgenic tobacco lines.

The A20/AN1 zinc-finger domain-containing proteins of the stress-associated proteins (SAPs) family are fast emerging as potential candidates for biotechnological approaches to improve abiotic stress tolerance in plants. We identified LmSAP, one of the SAPs genes in Lobularia maritima (L.) Desv., a halophyte brassicaceae, through its transcript accumulation in response to salinity and ionic stresses. Sequence homology analysis revealed that LmSAP contains two conserved zinc-finger domains A20 and AN1. Phylogeny analyses showed that LmSAP exhibited high amino acid sequence identity to other plant SAPs. Heterologous expression of LmSAP in yeast increased cell tolerance to salt and osmotic stress. In addition, the overexpression of LmSAP conferred high salt and ionic tolerance to transgenic tobacco plants. Transgenic tobacco seedlings showed higher survival rates and antioxidant activities under salt and ionic stresses. Enhanced antioxidant activities paralleled lower malondialdehyde and superoxide anion O2- levels in the LmSAP transgenic seedlings. Overall, our results suggest that overexpression of LmSAP enhanced salt tolerance by maintaining ionic balance and limiting oxidative and osmotic stresses.