Linalool Treatment Research Articles

Identification of Linalool Disaccharide Glycoside (Linalyl β-Vicianoside) in Soybean Leaves and Its Implication for Herbivore Resistance.

Linalool is anticipated to have significant ecological roles. In this study, linalyl 6-O-α-arabinopyranosyl-β-D-glucopyranoside (linalyl β-vicianoside: LinVic) was synthesized, and a linalool diglycoside purified from soybean leaves was identified as LinVic by using liquid chromatography-mass spectrometry. High levels of LinVic were detected in leaves and sepals during soybean plant growth. The LinVic content did not significantly increase following methyl jasmonate treatment of the leaves, indicating that its synthesis is independent of the jasmonic acid signaling pathway. In addition to LinVic, soybean also contains 1-octen-3-yl primeveroside. We treated soybean leaves with vaporized linalool and 1-octen-3-ol to determine whether the glycosylation system discriminates between these two volatile alcohols. Linalool treatment resulted in the accumulation of LinVic, while 1-octen-3-ol treatment caused little change in the amount of 1-octen-3-yl primeveroside, suggesting discrimination between these compounds. Linalool-treated soybean leaves exhibited increased resistance against common cutworms, indicating that LinVic may contribute to herbivore resistance.

Effects of linalool on respiratory neuron activity in the brainstem-spinal cord preparation from newborn rats.

Linalool and linalyl acetate are major components of lavender essential oil. These substances possess many biological activities, such as anti-inflammatory activity, analgesic and anxiolytic effects, and anticonvulsant properties, and they also induce modulation of neuronal activity in the autonomic nervous system. However, there are no reports of the direct effects of linalool on respiratory activity. In the present study, we analyzed the effects of linalool and linalyl acetate on central respiratory activity in the brainstem-spinal cord preparation isolated from newborn rats. Linalool dose-dependently decreased the rate of respiratory activity. This effect was reversed by bicuculline, suggesting that linalool enhanced inhibitory synaptic connections via GABAA receptors. In addition, linalool reduced the coefficient of variation of inspiratory burst intervals and thus could work to stabilize the respiratory rhythm. Linalyl acetate did not cause inhibitory effects as observed in linalool treatment. Linalool depressed burst activity of pre-inspiratory neurons in the medullary respiratory networks and increased the amplitude of inspiratory inhibitory postsynaptic potentials of pre-inspiratory neurons. We concluded that linalool caused inhibitory effects on respiratory rhythm generation mainly through activation of presynaptic GABAA receptors of pre-inspiratory neurons.

Volatile linalool activates grapevine resistance against downy mildew with changes in the leaf metabolome

Volatile organic compounds (VOCs) are produced by plants in response to biotic and abiotic stimuli. In grapevine, volatile terpenoids are triggered by downy mildew infection (caused by Plasmopara viticola), suggesting their involvement in plant defense responses. In particular, linalool was detected in leaves of downy mildew-resistant genotypes, but no information is available on its involvement in the defense mechanisms against P. viticola. The aim of this study was to investigate the defense mechanisms activated by linalool in grapevine leaves against P. viticola and to identify metabolic changes associated with linalool-induced resistance. Linalool treatment reduced downy mildew severity on leaf disks of susceptible grapevines (cultivar Pinot noir) and stimulated callose deposition at the sites of P. viticola infection. Moreover, the upregulation of defense-related genes was found in linalool-treated leaf disks, indicating the activation of grapevine defense mechanisms of salicylic acid and jasmonic acid pathways. Linalool treatment caused changes in the leaf metabolome of mock-inoculated and P. viticola-inoculated samples at one and six days post inoculation, as revealed by ultra-high pressure liquid chromatography-electrospray ionization-high-resolution quadrupole time of flight-mass spectrometry. Pathway analysis of annotated features with significant increases and decreases in abundance revealed the reprogramming of amino acid, phenylpropanoid, and terpenoid metabolisms in response to linalool treatment and P. viticola inoculation. In particular, features with significant increases in abundance in linalool-treated samples mainly belonged to putative phenylpropanoids, putative terpenoids, putative lipids, and lipid-like compounds, including molecules possibly associated with plant defense against pathogens, such as 2-phenylethanol, 2,4-heptadienal, α-terpineol, citral, and geraniol. These results demonstrated that linalool induces grapevine resistance against downy mildew, acting as a signaling molecule for plant resistance induction.

In vitro and in vivo anti-oomycetes activities and mechanisms of linalool against Saprolegnia ferax

Saprolegnia ferax (S. ferax) is one of the most highly pathogenic oomycetes species in freshwater aquatic environments causing serious saprolegniosis. Linalool is a kind of natural alternative of antibiotics existing in various plants essential oils. In this study, the in vitro and in vivo anti-oomycetes activity and mode of action of linalool against S. ferax CQT1 were investigated. For in vitro test, linalool exhibited mycelium growth and zoospore production inhibition at 0.1% (v/v, 0.862 mg/ml) and 0.025% (v/v, 0.216 mg/ml), respectively. The mycelium was wrinkled, deformed, and shrunken after treatment with linalool. Linalool altered membrane permeability, caused permanent damage, or membrane breakdown. Further, the transcriptomic analysis showed that: (1) Linalool affected the oxidation and metabolism of fatty acids, leading to mitochondrial dysfunction and the inhibition of energy supply. (2) The increase in glycerophospholipid metabolism is probably the cell response to deal with cellular damage. (3) The degradation of branched chain amino acids may inhibit amino acid metabolism and result in the dysfunction of protein synthesis. (4) ABC transporters contribute to metabolic resistance to linalool and efflux of linalool outside of cells. For in vivo test, the cumulative survival rate of grass carp treated with linalool was significantly higher than those untreated with linalool, moreover, microscopic histopathology analysis results showed that there was robust mucus cell density in fish skin, the dermis and epidermis were relatively intact after linalool treatment. In all, our findings would provide foundations for the potential use of linalool to prevent and control S. ferax infection in the aquaculture industry.

The Anti-Biofilm Potential of Linalool, a Major Compound from Hedychium larsenii, against Streptococcus pyogenes and Its Toxicity Assessment in Danio rerio.

The anti-biofilm and anti-virulence potential of the essential oil (E.O.) extracted from Hedychium larsenii M. Dan & Sathish was determined against Streptococcus pyogenes. A crystal violet assay was employed to quantify the biofilm. Linalool, a monoterpene alcohol from the E.O., showed concentration-dependent biofilm inhibition, with a maximum of 91% at a concentration of 0.004% (v/v). The AlamarBlueTM assay also confirmed Linalool's non-bactericidal anti-biofilm efficacy (0.004%). Linalool treatment impeded micro-colony formation, mature biofilm architecture, surface coverage, and biofilm thickness and impaired cell surface hydrophobicity and EPS production. Cysteine protease synthesis was quantified using the Azocasein assay, and Linalool treatment augmented its production. This suggests that Linalool destabilizes the biofilm matrix. It altered the expression of core regulons covRS, mga, srv, and ropB, and genes associated with virulence and biofilm formation, such as speB, dltA, slo, hasA, and ciaH, as revealed by qPCR analysis. Cytotoxicity analysis using human kidney cells (HEK) and the histopathological analysis in Danio rerio proved Linalool to be a druggable molecule against the biofilms formed by S. pyogenes. This is the first report on Linalool's anti-biofilm and anti-virulence potential against S. pyogenes.

Effects of linalool on Botrytis cinerea growth and control of tomato gray mold.

We examined the antifungal characteristics of linalool against Botrytis cinerea using plate inhibition assay and spore germination assay, and assessed the capacity of linalool in controlling tomato gray mold disease via tomato pot inoculation assay. The results showed that linalool exhibited strong inhibitive effects on mycelial growth of B. cinerea, with an EC50 value of 0.581 mL·L-1. In the spore germination test, linalool treatment inhibited spore germination in a dose-dependent manner. The electric conductivity and the malondialdehyde (MDA) contents were significantly increased in linalool-treated B. cinerea than that of the control, indicating that linalool induced oxidative damage and destroyed the cell membrane integrity in B. cinerea. The activities of the superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) in the linalool-treated B. cinerea were decreased significantly by 27.4%, 68.9% and 26.0%, respectively, suggesting that linalool inhibited the antioxidant activity of B. cinerea. In the pot experiment, the diameter of lesions in linalool-treated tomatoes was significantly smaller than that of the control. The activities of SOD, POD, CAT, polyphenol oxidase, and phenylalnine ammonialyase in the linalool-treated tomatoes increased, while the MDA content decreased, suggesting that linalool could alleviate the oxidative damage caused by B. cinerea and promote plant disease resistance. In summary, linalool had inhibitory effect on the growth of B. cinerea and could control gray mold disease in tomatoes. These findings could lay the foundation for developing bota-nical antifungal agents for management of tomato gray mold disease.

Revealing the Mechanisms for Linalool Antifungal Activity against Fusarium oxysporum and Its Efficient Control of Fusarium Wilt in Tomato Plants

Fusarium oxysporum f. sp. radicis-lycopersici (Forl) is a destructive soil-borne phytopathogenic fungus that causes Fusarium crown and root rot (FCRR) of tomato, leading to considerable field yield losses. In this study, we explored the antifungal capability of linalool, a natural plant volatile organic component, against Forl and its role in controlling FCRR symptoms in tomatoes. Our results showed that Forl mycelial growth was inhibited by the linalool treatment and that the linalool treatment damaged cell membrane integrity, enhanced reactive oxygen species levels, depleted glutathione, and reduced the activities of many antioxidant enzymes in Forl. Transcriptomic and proteomic analyses demonstrated that linalool also downregulated metabolic biosynthetic pathways at the transcript and protein levels, including redox, transporter activity, and carbohydrate metabolism in Forl. Moreover, linalool significantly decreased the expression of many Forl pathogenic genes, such as cell wall degrading enzymes (CWDEs) and G proteins, which is likely how a Forl infection was prevented. Importantly, exogenously applied linalool activated the salicylic acid (SA) and jasmonic acid (JA) defensive pathways to improve disease resistance and relieved the negative effects of Forl on plant growth. Taken together, we report that linalool is an effective fungicide against Forl and will be a promising green chemical agent for controlling FCRR.

Linalool controls the viability of Escherichia coli by regulating the synthesis and modification of lipopolysaccharide, the assembly of ribosome, and the expression of substrate transporting proteins

Escherichia coli (E. coli) is a Gram-negative bacterium and some pathogenic types may cause serious diseases, foods or food environments were the primary routes for its infection. Citrus aurantium L. var. amara Engl., a variety of sour orange, were used as a kind of non-conventional edible plant in China, but its antimicrobial activity and mechanisms were not well studied. Thus, in this study, EO from the flower of Citrus aurantium L. var. amara Engl. (CAEO) were studied as a kind of natural antimicrobial agent to control E. coli, our results showed that both of CAEO and its main component (linalool) exhibited strong antibacterial efficacy. Further, transcriptomic and proteomic analysis were carried out to explore cell response under linalool treatment and the main results included: (1) The synthesis and modification of lipopolysaccharide (LPS) was significantly influenced. (2) Ribosomal assembly and protein synthesis were significantly inhibited. (3) The expression of proteins related to the uptake of several essential substances was significantly changed. In all, our results would supply a theoretical basis for the proper use of CAEO and linalool as a promising antimicrobial agent to prevent and control E. coli infection in the future.

Linalool against Hafnia alvei, its antibacterial mechanism revealed by metabolomic analyses

Linalool, a major component of aromatic plant essential oils, has an antibacterial effect against Hafnia alvei, which can induce food spoilage. In this study, we aimed to investigate the antibacterial effect and mechanism of linalool against H. alvei. Linalool treatment resulted in excessive reactive oxygen species (ROS) accumulation, which resulted in destruction of the cell membrane. According to the field emission scanning electron microscopy (FESEM) and flow cytometry (FCM) results, it was confirmed that the membrane structure of H. alvei was destroyed. Moreover, metabolomics analysis showed that there were 64 and 73 differentially expressed metabolites screened by mass spectrometry in positive and negative ion modes, respectively, suggesting that nucleic acid metabolism, the respiratory chain and energy metabolism were negatively affected by linalool. Biochemical validation confirmed that the activities of malate dehydrogenase (MDH), and succinate dehydrogenase (SDH) and respiratory metabolism were reduced after linalool treatment. It was therefore concluded that the cell membrane was damaged, nucleic acid metabolism was disturbed, and energy metabolism was reduced by inhibition of the respiratory chain and tricarboxylic acid (TCA) cycle. These results revealed the antibacterial mechanism of linalool against H. alvei, and support the use of linalool as a potential natural preservative to control H. alvei.

Alleviating Effects of Linalool Fumigation on Botrytis cinerea Infections in Postharvest Tomato Fruits

Gray mold is a disease that often occurs in postharvest tomato fruits, leading to a significant decline in the fruits’ quality. In production, pesticides are mostly used to control gray mold, which negatively affect both the environment and food safety. The purpose of this article is to study the effects of linalool fumigation on controlling gray mold in tomato fruits and to further investigate the mechanism of linalool function, so as to provide technical support and a theoretical basis for the application of linalool in the green control of tomato gray mold. The results of the in vitro experiments showed that linalool fumigation had a strong inhibitory effect on the mycelial growth of Botrytis cinerea and that the fumigation of linalool inhibited the expansion of pathogens on tomato fruits. The disease index of tomato fruits in the linalool treatment was always lower than that of the control within 72 h after inoculation with Botrytis cinerea; at the end of experiment, the disease index of the control reached 100.0, which was only 8.0 with the 30 μL/L linalool treatment. Linalool fumigation increased the antioxidant capacity of the tomato fruits under Botrytis cinerea infection through regulating activities of SOD, POD and CAT as well as the ascorbic acid (AsA) content, which could be responsible for the lower malondialdehyde (MDA) accumulation. Linalool fumigation increased the activities of polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) in tomato fruits, indicating that secondary metabolism was involved in the function of linalool in the regulation of tomato fruit resistance to pathogens. As the main enzymes related to cell structure, polygalacturonase (PG), cellulase (CL) and β-galactosidase (β-GAL) were inhibited by linalool, which could protect cell wall structures from damage, and strengthened the mechanical barrier against pathogen access to fruit flesh.

Mechanisms underlying the inhibitory effects of linalool on Aspergillus flavus spore germination.

Biogenic volatile organic compounds hold remarkable potential for controlling fungal decay in agro- and food products. Recently, we reported that linalool, the major volatile component of the Zanthoxylum schinifolium pericarp, showed great potential as a biofumigant to control Aspergillus flavus growth in postharvest grains. In this study, the inhibitory effects of linalool on A. flavus growth in stored grains and its underlying mechanism were investigated through transcriptomic and biochemical analyses. Linalool vapor at 800 μL/L can effectively prevent A. flavus growth in 22% moisture wheat grains. Linalool at 2 μL/mL completely inhibited the germination of A. flavus spores, and 10 μL/mL caused spore death. Scanning electron microscopy revealed that linalool treatment caused wrinkling and spore breakage. Transcriptomics showed that 3806 genes were significantly differentially expressed in A. flavus spores exposed to 2 μL/mL linalool, predominantly showing enrichment regarding the ribosome, DNA replication, glutathione metabolism, peroxisome, and MAPK signaling pathways. Flow cytometry showed that linalool treatment caused hyperpolarization of mitochondrial membrane potential. 4,6-Diamidino-2-phenylindole staining indicated that linalool caused DNA fragmentation in A. flavus spores, and monodansylcadaverine staining confirmed that linalool induced autophagy in A. flavus spores. We thus propose that linalool can damage the plasma membrane, cause mitochondrial dysfunction and DNA damage, and induce autophagy in A. flavus spores. These findings considerably improve our understanding of the mechanisms underlying the inhibitory effects of linalool on A. flavus, which is crucial regarding the development of applications to prevent postharvest grain spoilage due to A. flavus infestations. KEY POINTS: • The inhibitory potency of linalool on A. flavus spore germination was determined. • Transcriptomic analyses were performed to identify differentially expressed genes of A. flavus exposed to linalool. • A functional mechanism underlying the inhibitory effects of linalool on A. flavus spore germination is proposed.

Antibacterial Mechanism of Linalool against Pseudomonas fragi: A Transcriptomic Study.

Pseudomonas fragi is the dominant spoilage bacterium that causes the deterioration of chilled meat. Our previous study showed that linalool has potent antibacterial activity against P. fragi, but its antibacterial mechanism is unclear. To explore the antibacterial mechanism of linalool against P. fragi, this study used RNA-seq technology to perform transcriptome analysis of P. fragi samples with or without linalool treatment (1.5 mL/L) for 2 h. The results showed that linalool treatment disrupted the extracellular lipopolysaccharide synthesis pathway in P. fragi and activated fatty acid metabolism and ribosomal function to compensate for cell membrane damage. The energy metabolism of P. fragi was severely disturbed by linalool, and multiple ATP synthases and ATP transportases were overexpressed in the cells but could not guarantee the consumption of ATP. The simultaneous overexpression of multiple ribosomal functional proteins and transporters may also place an additional burden on cells and cause them to collapse.

Linalool Activates Oxidative and Calcium Burst and CAM3-ACA8 Participates in Calcium Recovery in Arabidopsis Leaves.

Plants produce linalool to respond to biotic stress, but the linalool-induced early signal remains unclear. In wild-type Arabidopsis, plant resistance to diamondback moth (Plutella xylostella) increased more strongly in a linalool-treated group than in an untreated control group. H2O2 and Ca2+, two important early signals that participated in biotic stress, burst after being treated with linalool in Arabidopsis mesophyll cells. Linalool treatment increased H2O2 and intracellular calcium concentrations in mesophyll cells, observed using a confocal microscope with laser scanning, and H2O2 signaling functions upstream of Ca2+ signaling by using inhibitors and mutants. Ca2+ efflux was detected using non-invasive micro-test technology (NMT), and Ca2+ efflux was also inhibited by NADPH oxidase inhibitor DPI (diphenyleneiodonium chloride) and in cells of the NADPH oxidase mutant rbohd. To restore intracellular calcium levels, Ca2+-ATPase was activated, and calmodulin 3 (CAM3) participated in Ca2+-ATPase activation. This result is consistent with the interaction between CAM7 and Ca2+-ATPase isoform 8 (ACA8). In addition, a yeast two-hybrid assay, firefly luciferase complementation imaging assay, and an in vitro pulldown assay showed that CAM3 interacts with the N-terminus of ACA8, and qRT-PCR showed that some JA-related genes and defense genes expressions were enhanced when treated with linalool in Arabidopsis leaves. This study reveals that linalool enhances H2O2 and intracellular calcium concentrations in Arabidopsis mesophyll cells; CAM3-ACA8 reduces intracellular calcium concentrations, allowing cells to resume their resting state. Additionally, JA-related genes and defense genes’ expression may enhance plants’ defense when treated with linalool.

Linalool, the main volatile constituent from Zanthoxylum schinifolium pericarp, prevents growth of Aspergillus flavus in post-harvest grains

Plant volatile organic compounds (PVOCs) have shown great potential as alternatives to synthetic insecticides or fungicides for stored grain management. Zanthoxylum schinifolium pericarp, a traditional Chinese spice, is conventionally buried in grain piles to prevent fungal spoilage of stored grains in China. However, the chemical basis and antifungal mechanism of PVOCs from Z. schinifolium pericarp by which they inhibit spoilage remain unclear. In this study, the effectiveness of PVOCs from Z. schinifolium pericarp against Aspergillus flavus growth in high-moisture wheat grains was studied under simulated storage conditions. The growth of A. flavus in the grains was inhibited in a dose-dependent manner. The chemical composition of PVOCs from Z. schinifolium pericarp was determined using gas chromatography-mass spectrometry; linalool (50.31%) and d-limonene (20.92%) were the two main components. An antifungal experiment showed that 0.571 and 1.2 μL/mL concentrations of linalool completely inhibited A. flavus growth upon vapor and liquid contact, respectively. d-limonene showed much lower antifungal activity, indicating that linalool is responsible for the antifungal activity of PVOCs from Z. schinifolium pericarp. Linalool treatment disrupted the cell membrane of A. flavus, resulting in increased electrolyte leakage and A260nm in the culture supernatant. After 6 h of exposure to 1.2 μL/mL linalool, metabolomic analysis revealed 90 differentially expressed metabolites in A. flavus mycelia, including 69 upregulated and 21 downregulated metabolites. It was speculated that linalool inhibited A. flavus by disrupting the permeability and integrity of cell membranes, tricarboxylic acid cycle, and ATP binding cassette transport and by inducing mitochondrial dysfunction and oxidative stress. This study shows the potential of PVOCs from Z. schinifolium pericarp as biofumigants for stored grain management and provides a new perspective on their antifungal mechanism against A. flavus growth.

Robust behavioural effects in response to acute, but not repeated, terpene administration in Zebrafish (Danio rerio)

Terpenes are fragrant aromatic compounds produced by a variety of plants, most notably cannabis and hops. With increasing legalization of cannabis there is a need to better understand the behavioural effects of terpenes and ultimately their therapeutic value. Our study investigated the dose-dependent impact of three terpenes (limonene 0.25, 0.5, 0.75%; β-myrcene 0.001, 0.01, 0.1%; and 0.0001, 0.001, 0.00125% linalool) on zebrafish (Danio rerio) behaviour when exposed both acutely and repeatedly over a 7-day period. Anxiety-like behaviour, boldness, and locomotion were assessed using the open field test and the novel object approach test. In the acute dosing experiment, limonene and β-myrcene exposed groups demonstrated a significant decrease in locomotion, a decrease in anxiety-like behaviour, and an increase in boldness, while linalool treatment groups demonstrated only minor alterations in locomotion. Moreover, repeated exposure to limonene (0.39%) or β-myrcene (0.0083%) for a seven day period did not result in any significant behavioural effects. In conclusion, our study provides support for an anxiolytic and sedative effect in zebrafish in response to acute limonene and β-myrcene exposure that is no longer present after one week of repeated exposure.

Linalool Inhibits MCF-7 Tumor Growth in a Xenograft Model by Apoptosis Induction and Immune Modulation

In this study, the anti-cancer activity of linalool was investigated in MCF-7 breast cancer-bearing mice. Natural killer (NK) and B cell populations in peripheral blood were studied by flow cytometry. The expressions of proliferating cell nuclear antigen (PCNA) and Ki-67 in xenograft tumors were evaluated by immunohistochemistry. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to investigate apoptosis induction in an in vivo model. The results indicated that linalool possesses an inhibitory effect on breast cancer growth in the xenograft model. Linalool reduced B cell counts, but increased NK cell counts in mice peripheral blood. The immunosignals of PCNA and Ki-67 were significantly lower in the linalool treatment group than those of the control group. The TUNEL assay showed that linalool significantly induced apoptosis compared to the control group. The findings of this study provide insight and evidence on the antiproliferative activity of linalool on human breast cancer.

Combinatorial Antimicrobial Efficacy and Mechanism of Linalool Against Clinically Relevant Klebsiella pneumoniae

Antibiotic–adjuvant combinatory therapy serves as a viable treatment option in addressing antibiotic resistance in the clinical setting. This study was carried out to assess and characterize the adjuvant potential and mode of action of linalool against carbapenemase-producing Klebsiella pneumoniae (KPC-KP). Linalool exhibited bactericidal activity alone (11,250 μg/ml) and in combination with meropenem (5,625 μg/ml). Comparative proteomic analysis showed significant reduction in the number of cytoplasmic and membrane proteins, indicating membrane damage in linalool-treated KPC-KP cells. Upregulation of oxidative stress regulator proteins and downregulation of oxidative stress-sensitive proteins indicated oxidative stress. Zeta potential measurement and outer membrane permeability assay revealed that linalool increases the bacterial surface charge as well as the membrane permeability. Intracellular leakage of nucleic acid and proteins was detected upon linalool treatment. Scanning and transmission electron microscopies further revealed the breakage of bacterial membrane and loss of intracellular materials. Linalool induced oxidative stress by generating reactive oxygen species (ROS) which initiates lipid peroxidation, leading to damage of the bacterial membrane. This leads to intracellular leakage, eventually killing the KPC-KP cells. Our study demonstrated that linalool possesses great potential in future clinical applications as an adjuvant along with existing antibiotics attributed to their ability in disrupting the bacterial membrane by inducing oxidative stress. This facilitates the uptake of antibiotics into the bacterial cells, enhancing bacterial killing.

Antimicrobial Activity and Proposed Action Mechanism of Linalool Against Pseudomonas fluorescens.

In this study, linalool, one of the principal components of essential oils, was used as an antibacterial agent to investigate the antibacterial activity and mechanism of linalool against Pseudomonas fluorescens. The reduction in membrane potential (MP), leakage of alkaline phosphatase (AKP) and the release of macromolecules, including DNA, RNA and protein confirmed that damage to cell wall membrane structure and leakage of cytoplasmic contents were due to the linalool treatment. Furthermore, the decrease of enzyme activity, including the succinate dehydrogenase (SDH), malate dehydrogenase (MDH), pyruvate kinase (PK), and ATPase indicated that linalool could lead to metabolic dysfunction and inhibit energy synthesis. In addition, the activity of respiratory chain dehydrogenase and metabolic activity of respiration indicated that linalool inhibits cellular respiration. These results revealed that linalool had strong antibacterial activity against P. fluorescens via membrane damage, bacterial metabolic and oxidative respiratory perturbations, interfering in cellular functions and even causing cell death. It was suggested that linalool may be a new potential source as food antiseptics in food systems.

Antibacterial Activity and Mechanism of Linalool against Shewanella putrefaciens.

The demand for reduced chemical preservative usage is currently growing, and natural preservatives are being developed to protect seafood. With its excellent antibacterial properties, linalool has been utilized widely in industries. However, its antibacterial mechanisms remain poorly studied. Here, untargeted metabolomics was applied to explore the mechanism of Shewanella putrefaciens cells treated with linalool. Results showed that linalool exhibited remarkable antibacterial activity against S. putrefaciens, with 1.5 µL/mL minimum inhibitory concentration (MIC). The growth of S. putrefaciens was suppressed completely at 1/2 MIC and 1 MIC levels. Linalool treatment reduced the membrane potential (MP); caused the leakage of alkaline phosphatase (AKP); and released the DNA, RNA, and proteins of S. putrefaciens, thus destroying the cell structure and expelling the cytoplasmic content. A total of 170 differential metabolites (DMs) were screened using metabolomics analysis, among which 81 species were upregulated and 89 species were downregulated after linalool treatment. These DMs are closely related to the tricarboxylic acid (TCA) cycle, glycolysis, amino acid metabolism, pantothenate and CoA biosynthesis, aminoacyl-tRNA biosynthesis, and glycerophospholipid metabolism. In addition, linalool substantially affected the activity of key enzymes, such as succinate dehydrogenase (SDH), pyruvate kinase (PK), ATPase, and respiratory chain dehydrogenase. The results provided some insights into the antibacterial mechanism of linalool against S. putrefaciens and are important for the development and application of linalool in seafood preservation.

Linalool Induces Reactive Oxygen Species Mediated Apoptosis in Human Oral Squamous Carcinoma Cells

Development of oral cancer is associated with excessive cell proliferation, deregulation in cellular differentiation, insufficient apoptosis and genomic instability. Linalool is a natural monoterpene, contained in edible plants, considered to have potent antioxidant nature. In this context, it should be necessitate in broadening the understanding of the anticancer potential of linalool in experimental oral cancer model, which could be beneficial in anticancer therapy. Hence, this study is intended to assess the anticancer quality of linalool in keratin-forming tumor cell line. 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide assay was conducted to find out the half maximal inhibitory concentration of linalool in keratinforming tumor cell line. Linalool induced the loss of cell viability or cell death by a clear dose and time dependent manner. The half maximal inhibitory concentration on keratin-forming tumor cell line cells is 13, 24 and 45 μM for 24, 48 and 72 h respectively. Morphological changes, increased level of intracellular level of reactive oxygen species and loss of mitochondrial transmembrane potential of keratin-forming tumor cell line cells were evidently visualized on staining. Early growth 1 phase arrest in the cell cycle of keratin-forming tumor cell line cells was confirmed on dose and time dependent manner when compared with control during linalool treatment. Linalool treatment in various concentrations in vitro in keratinforming tumor cell line cells was able to reduce cell viability as compared to its counterparts. This study showed that linalool was a potent growth inhibitor of keratin-forming tumor cell line. In near future in vivo screening for its anticancer activity will be warranted and a linkage map will be developed between in vivo and in vitro studies.