Hormesis management of Moringa oleifera with exogenous application of plant growth regulators under saline conditions.

The study investigated the adaptability of Moringa oleifera to saline conditions, focusing on its hormesis behavior. It also examined how various plant growth regulators affected growth, physiological parameters, and bioactive compounds of moringa. In the first phase, different NaCl stress levels (0, 50, 100, 150, 200, and 250 mM) were applied. Notably, significant stimulation was observed at 100 mM stress for growth, total phenolics, total flavonoids and total chlorophyll content while 150 mM stress had a marked inhibitory effect, with survival decreasing at 200 and 250 mM NaCl levels. A 38% reduction in root attributes and shoot length, along with a 55% decrease in leaf score, was observed at 150 mM stress. Total phenolics showed a positive correlation with growth attributes. In the second phase, moringa plants grown under 50, 100, and 150 mM NaCl stress were treated with various plant growth regulators, including cytokinin (50 mg L-1), thiourea (5 mM), bezyl amino purine (BAP @50 mg L-1), salicylic acid (50 mg L-1), hydrogen peroxide (H2O2@120 µM), or ascorbic acid (50 mg L-1) to mitigate adverse effects of salinity. Cytokinin, BAP, and salicylic acid applications improved salinity tolerance, enhancing enzymatic, and non-enzymatic antioxidants, and the abundance of kaempferol, quercetin, hydroxybenzoic, and hydroxycinnamic acids. Pearson correlation and principal component analysis manifested relationships among growth parameters, antioxidant activities, flavonoids, and phenolic acids. This study provides new insights into hormesis management for moringa plants and the influence of plant growth regulators on flavonoids and phenolic acid levels in moringa leaves under saline conditions.

This study represents the first exploration of hormesis management in Moringa oleifera dual influence of changing soil conditions and foliar application of plant growth regulators. Additionally, this research fills a gap examining the variations in flavonoids (kaempferol and quercetin), hydroxycinnamic acids and hydroxybenzoic acids in moringa leaves concerning varying salinity levels and the exogenous application of plant growth regulators. Further, the study underscores the correlation among secondary metabolites, antioxidant activities and plant growth behavior.

Structural variation underlies functional diversity at methyl salicylate loci in tomato.

Methyl salicylate is an important inter- and intra-plant signaling molecule, but is deemed undesirable by humans when it accumulates to high levels in ripe fruits. Balancing the tradeoff between consumer satisfaction and overall plant health is challenging as the mechanisms regulating volatile levels have not yet been fully elucidated. In this study, we investigated the accumulation of methyl salicylate in ripe fruits of tomatoes that belong to the red-fruited clade. We determine the genetic diversity and the interaction of four known loci controlling methyl salicylate levels in ripe fruits. In addition to Non-Smoky Glucosyl Transferase 1 (NSGT1), we uncovered extensive genome structural variation (SV) at the Methylesterase (MES) locus. This locus contains four tandemly duplicated Methylesterase genes and genome sequence investigations at the locus identified nine distinct haplotypes. Based on gene expression and results from biparental crosses, functional and non-functional haplotypes for MES were identified. The combination of the non-functional MES haplotype 2 and the non-functional NSGT1 haplotype IV or V in a GWAS panel showed high methyl salicylate levels in ripe fruits, particularly in accessions from Ecuador, demonstrating a strong interaction between these two loci and suggesting an ecological advantage. The genetic variation at the other two known loci, Salicylic Acid Methyl Transferase 1 (SAMT1) and tomato UDP Glycosyl Transferase 5 (SlUGT5), did not explain volatile variation in the red-fruited tomato germplasm, suggesting a minor role in methyl salicylate production in red-fruited tomato. Lastly, we found that most heirloom and modern tomato accessions carried a functional MES and a non-functional NSGT1 haplotype, ensuring acceptable levels of methyl salicylate in fruits. Yet, future selection of the functional NSGT1 allele could potentially improve flavor in the modern germplasm.

New trends in methyl salicylate sensing and their implications in agriculture.

Methyl salicylate (MeSal) is an organic compound present in plants during stress events and is therefore a key marker for early plant disease detection. It has usually been detected by conventional methods that require bulky and costly equipment, such as gas chromatography or mass spectrometry. Currently, however, chemical sensors provide an alternative for MeSal monitoring, showing good performance for its determination in the vapour or liquid phase. The most promising concepts used in MeSal determination include sensors based on electrochemical and conductometric principles, although other technologies based on mass-sensitive, microwave, or spectrophotometric principles also show promise. The receptor elements or sensitive materials are shown to be part of the key elements in these sensing technologies. A literature survey identified a significant contribution of bioreceptors, including enzymes, odourant-binding proteins or peptides, as well as receptors based on polymers or inorganic materials in MeSal determination. This work reviews these concepts and materials and discusses their future prospects and limitations for application in plant health monitoring.

25-Year Storage of Human Choroid Plexus in Methyl Salicylate Preserves Its Antigen Immunoreactivity.

OBJECTIVE: Immunohistochemical investigation of archival histological material is a serious problem, since long-term storage of biological tissues, most often in formalin, leads to a loss of antigenic properties. However, the biological material can also be stored in the clearing agent methyl salicylate. The aim of this study was to assess the antigenicity of the human choroid plexus after extra long-term storage in methyl salicylate. MATERIAL AND METHOD: The study was performed on samples of fixed human choroid plexus (occasionally with attached neighboring pineal gland) stored in either methyl salicylate or paraffin blocks for 25 years. Chromogenic and fluorescence immunohistochemistry of vimentin, GFAP, type IV collagen, ß-catenin, α-smooth muscle actin, von Willebrand factor, CD68, mast cell tryptase, TMEM119, and synaptophysin was carried out. RESULTS: The storage of human choroid plexus in methyl salicylate for 25 years does not impair its histomorphology and preserves the properties of all the antigens assessed, which makes their immunohistochemical visualization possible using both light and fluorescence microscopy. Additionally, we found that long-term storage of human choroid plexus in methyl salicylate does not cause an increase in autofluorescence. CONCLUSION: Methyl salicylate can be recommended as a medium for long-term storage of biological tissue, as it provides excellent brain tissue preservation and retains its antigenic properties for up to 25 years.

Identification of Salicylates in Willow Bark (Salix Cortex) for Targeting Peripheral Inflammation.

Salix cortex-containing medicine is used against pain conditions, fever, headaches, and inflammation, which are partly mediated via arachidonic acid-derived prostaglandins (PGs). We used an activity-guided fractionation strategy, followed by structure elucidation experiments using LC-MS/MS, CD-spectroscopy, and 1D/2D NMR techniques, to identify the compounds relevant for the inhibition of PGE2 release from activated human peripheral blood mononuclear cells. Subsequent compound purification by means of preparative and semipreparative HPLC revealed 2'-O-acetylsalicortin (1), 3'-O-acetylsalicortin (2), 2'-O-acetylsalicin (3), 2',6'-O-diacetylsalicortin (4), lasiandrin (5), tremulacin (6), and cinnamrutinose A (7). In contrast to 3 and 7, compounds 1, 2, 4, 5, and 6 showed inhibitory activity against PGE2 release with different potencies. Polyphenols were not relevant for the bioactivity of the Salix extract but salicylates, which degrade to, e.g., catechol, salicylic acid, salicin, and/or 1-hydroxy-6-oxo-2-cycohexenecarboxylate. Inflammation presents an important therapeutic target for pharmacological interventions; thus, the identification of relevant key drugs in Salix could provide new prospects for the improvement and standardization of existing clinical medicine.

Effects of preheating and drying methods on pyridoxine, phenolic compounds, ginkgolic acids, and antioxidant capacity of Ginkgo biloba nuts.

Although ginkgo nuts are very nutritious and loaded with numerous bioactive compounds, the nuts contain significant levels of unwanted compounds (ginkolic acids) which are toxic to consumption. To reduce or eliminate these toxic compounds without impacting the nutritional value and the bioactivity of the final product, an appropriate processing technology is needed. Thus, the effect of preheating (90 and 120°C) prior to drying (freeze drying: FD, hot air drying: HAD, and HAD in tandem with FD: HAD-FD) was evaluated on ginkgolic acids, pyridoxine analogues, phenolic compounds, and antioxidant properties of ginkgo nuts. Our results pointed out a significant decrease (below 50%) of ginkgolic acids in ginkgo nuts samples processed at 90°C compared to the control. The major compounds found after treatments were respectively, kaempferol (36.66-354.38 µg/g), quercetin (9.04-183.71 µg/g), and caffeic acid (19.66-106.88 µg/g). Principal component analysis (PCA) revealed that preheating at 90°C prior to HAD-FD would be a proper and reasonable approach for preserving the bioactive compounds and antioxidant capacity of ginkgo nuts (EC50 ranged from 2.25 to 4.60 mg/mL) while significantly reducing their content in toxic compounds.

Methyl salicylate, a grape and wine chemical marker and sensory contributor in wines elaborated from grapes affected or not by cryptogamic diseases.

Methyl salicylate (MeSA) is a plant metabolite that induces plant defence resistance and an odorous volatile compound presenting green nuances. This volatile compound was shown to be present in wine samples, sometimes at concentrations above its olfactory detection threshold. MeSA is localized in grapes, particularly in the skins and stems, and is extracted during red wine vinification. It was detected at the highest concentrations in wines of several grape varieties, made from grapes affected by cryptogamic diseases, namely downy mildew caused by Plasmopara viticola, and black rot caused by Guignardia bidwellii. It has also been detected in wines from vines affected by Esca, a Grapevine Trunk Disease. MeSA can also be considered to be a chemical marker in grapes and wine indicative of the level of development of several vine cryptogamic diseases.

Effect of Leaf Maturity on Host Habitat Location by the Egg-Larval Parasitoid Ascogaster reticulata.

Adoxophyes honmai, a serious pest of tea plants, prefers to lay eggs on mature tea leaves rather than young leaves. Here, we examined a hypothesis that Ascogaster reticulata, an egg-larval parasitoid of A. honmai, increases the likelihood of encountering host egg masses by searching mature tea leaves when host-derived cues are not available. In a dual-choice bioassay using a four-arm olfactometer, A. reticulata preferred odor from intact, mature leaves versus young leaves. Based on volatile analysis with gas chromatography-mass spectrometry (GC-MS), we identified 5 and 10 compounds from mature and young leaf volatiles, respectively. The 5 components in the extract from intact mature leaves included (Z)-3-hexenyl acetate, (E)-ß-ocimene, linalool, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), and methyl salicylate. When each individual compound, or quaternary and quintenary blends of them, ratios of which were adjusted to match those of mature leaf volatiles, were provided, parasitoids preferred the full mixture and the quaternary blend devoid of DMNT to the solvent control. Methyl salicylate, one of the components of preferred blends, was not detected among young leaf volatiles. We concluded that the volatile composition of tea leaves changes, depending on their maturity, and that this composition affects foraging behavior of the parasitoid, which is closely related to the host herbivore's oviposition preference.

Submerged fermentation of Ginkgo biloba seed powder using Eurotium cristatum for the development of ginkgo seeds fermented products.

BACKGROUND: Ginkgo biloba seeds are well known for the significant curative effects on relieving cough and asthma. However, the development of products from ginkgo seeds still falls behind at present, resulting in a great waste of ginkgo seeds' resource. In this work, submerged fermentation of ginkgo seed powder using Eurotium cristatum was studied to investigate its feasibility as a new processing method. RESULTS: To promote the growth of E. cristatum, the optimum fermentation medium was 80.0 g L-1 of ginkgo seed powder with addition of 5.0 g L-1 calcium chloride (CaCl2 ), 4.0 g L-1 magnesium sulfate (MgSO4 ), 1.25 g L-1 zinc sulfate (ZnSO4 ) and 0.65 g L-1 iron(II) sulfate (FeSO4 ). The optimum fermentation conditions were pH 5.8 ± 0.1, inoculum size 5.1 × 106 CFU mL-1 , liquid medium volume 100 mL in 250-mL Erlenmeyer flask and fermentation 4 days. Through fermentation, the production of lovastatin in fermentation broth could reach up to 32.97 ± 0.17 µg mL-1 and the total antioxidant capacity was improved by more than two-fold. In addition, 40.15% of the ginkgotoxin in ginkgo seed powder was degraded while the entire degradation of ginkgolic acids was obtained. Moreover, fermented ginkgo seed powder suspension presented pleasant fragrances, and the activities of amylase and protease were enhanced to 11.30 ± 0.10 U mL-1 and 23.01 ± 0.20 U mL-1 , respectively. CONCLUSIONS: Submerged fermentation using E. cristatum could significantly enhance the functional value and safety of ginkgo seed powder, and had great potential to become a novel processing method for the development of ginkgo seeds fermented products. © 2020 Society of Chemical Industry.

Gaultheria longibracteolata, an alternative source of wintergreen oil.

Gaultheria longibracteolata (Ericaceae) has been traditionally used by different linguistic groups in Yunnan Province, China, but it has not been well studied. Through our ethnobotanical study in Lüchun County of Yunnan, we found that this species has multiple traditional uses including food, medicine, and worship. The essential oils from the root, stem, and leaf were investigated by both GC-MS and anti-bacterial assays. The GC-MS study showed that methyl salicylate is the main (>90%) component of the essential oil, and the oil composition extracted from different plant parts showed some similarities. The oil of G. longibracteolata displayed antimicrobial activity against Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus, which is likely due to its methyl salicylate content. Gautheria longbracteolata appears to be a useful natural wintergreen oil substitute, but further studies are needed to develop this product.

Engineered Bacterial Production of Volatile Methyl Salicylate.

The engineering of microbial metabolic pathways over the last two decades has led to numerous examples of cell factories used for the production of small molecules. These molecules have an array of utility in commercial industries and as in situ expressed biomarkers or therapeutics in microbial applications. While most efforts have focused on the production of molecules in the liquid phase, there has been increasing interest in harnessing microbes' inherent ability to generate volatile compounds. Here, we optimized and characterized the production of methyl salicylate, an aromatic compound found mainly in plants, using a common lab strain of E. coli. We utilized genetic components from both microbes and plants to construct the volatile metabolite circuit cassette. In order to maximize production, we explored expression of methyl salicylate precursors, upregulation of expression by increasing ribosomal binding strength and codon optimization of the methyl transferase gene obtained from plant Petunia x hybrida. Last, we validated and quantified the production of methyl salicylate with liquid chromatography or gas chromatography mass spectrometry (LC-MS or GC-MS) and found that the codon optimized strain with precursor supplementation yielded the highest production compared to the other strains. This work characterizes an optimized metabolite producing genetic circuit and sets the stage for creation of an engineered bacteria diagnostic to be used in volatile assays.

Effect of different drying methods on product quality, bioactive and toxic components of Ginkgo biloba L. seed.

BACKGROUND: Ginkgo biloba seeds are used as a functional food across Asia. However, the presence of toxic compounds has limited their application. In this study, freeze drying, infrared drying, hot-air drying and pulsed-vacuum drying were used to dry G. biloba seeds. A comprehensive analysis was performed on their product quality, antioxidant activities, bioactive and toxic components. RESULTS: Results showed that the drying methods had a significant influence on product quality with freeze drying being superior due to the minimal microstructural damage, followed by infrared drying and pulsed-vacuum drying. Infrared-dried product possessed the strongest antioxidant activities and higher bioactive compound content than hot-air-dried and pulsed-vacuum-dried product. Toxic compounds in fresh G. biloba seeds (ginkgotoxin, ginkgolic acid and cyanide) were reduced markedly by drying. Ginkgotoxin was reduced fourfold, and the contents of acrylamide, ginkgolic acid and cyanide in dried G. biloba seeds were reduced to the scope of safety. Amongst the four drying methods, infrared drying had the shortest drying time, and its product showed higher quality and bioactive compound content, and stronger antioxidant activities. CONCLUSIONS: These findings will offer salient information for selecting a drying method during the processing of ginkgo seeds. Infrared drying could be considered as a multiple-effect drying method in the processing of ginkgo seeds. © 2020 Society of Chemical Industry.

A controlled cross-over study to evaluate the efficacy of improvised dry and wet emergency decontamination protocols for chemical incidents.

The UK Initial Operational Response (IOR) to chemical incidents includes improvised decontamination procedures, which use readily available materials to rapidly reduce risk to potentially exposed persons. A controlled, cross-over human volunteer study was conducted to investigate the effectiveness of improvised dry and wet decontamination procedures on skin, both alone, and in sequence. A simulant contaminant, methyl salicylate (MeS) in vegetable oil with a fluorophore was applied to three locations (shoulder, leg, arm). Participants then received no decontamination (control) or attempted to remove the simulant using one of three improvised protocols (dry decontamination; wet decontamination; combined dry and wet decontamination). Simulant remaining on the skin following decontamination was quantified using both Gas Chromatography Tandem Mass Spectrometry (GC-MSMS) for analysis of MeS and UV imaging to detect fluorophores. Additionally, urine samples were collected for 24 hours following application for analysis of MeS. Significantly less simulant was recovered from skin following each improvised decontamination protocol, compared to the no decontamination control. Further, combined dry and wet decontamination resulted in lower recovery of simulant when compared to either dry or wet decontamination alone. Irrespective of decontamination protocol, significantly more simulant remained on the shoulders compared to either the arms or legs, suggesting that improvised decontamination procedures are less effective for difficult to reach areas of the body. There was no effect of decontamination on excreted MeS in urine over 24 hours. Overall, findings indicate that improvised decontamination is an effective means of rapidly removing contaminants from skin, and combinations of improvised approaches can increase effectiveness in the early stages of decontamination and in the absence of specialist resources at an incident scene. However, the variable control and consistency of improvised decontamination techniques means that further intervention is likely to be needed, particularly for less accessible areas of the body.

Intake of Dietary Salicylates from Herbs and Spices among Adult Polish Omnivores and Vegans.

There is a growing body of evidence that a diet rich in bioactive compounds from herbs and spices has the ability to reduce the risk of chronic diseases. The consumption of herbs and spices is often overlooked in the studies on food intake. However, measurement of dietary intake of these products, as a source of bioactive compounds, including salicylates, has recently gained much significance. The aims of the study were (i) to assess the intake of herbs and spices at the individual level and (ii) to calculate the dietary salicylates intake from herbs and spices among adult omnivores and vegans. The study group consisted of 270 adults aged 19 to 67 years, including 205 women and 65 men. Among all, 208 individuals were following an omnivorous diet while 62 were vegans. A semi-quantitative food frequency questionnaire (FFQ) was designed to assess the habitual intake of 61 fresh and dried herbs and spices during the preceding three months. The five most frequently eaten herbs among omnivores were parsley, garlic, dill, marjoram and basil, while among vegans they were garlic, parsley, ginger, basil and dill. An average intake of all condiments included in the study was 22.4 ± 18.4 g/day and 25.8 ± 25.9 g/day for both omnivores and vegans, respectively (p = 0.007). Estimated medial salicylates intake was significantly higher among vegans (p = 0.000) and reached 5.82 mg/week vs. 3.13 mg/week for omnivores. Our study confirmed that herbs and spices are important sources of salicylates; however, the type of diet influenced their level in the diet. Vegans consume significantly more total salicylates than omnivores.

Sex Attractant Pheromone of the Japanese Slave-Making Ant, Polyergus samurai.

The objective of our project was to identify the sex attractant pheromone of queens of the slave-making ant Polyergus samurai, which was characterized as a blend of methyl 6-methylsalicylate and methyl 3-ethyl-4-methylpentanoate. Both compounds were identified in volatiles collected from squashed heads of queens, and in field trials, a blend of the two compounds was highly attractive to males. In contrast, males were not attracted to a blend of methyl 6-methylsalicylate with 3-ethyl-4-methylpentanol. Thus, the pheromone blend of this species is analogous to that of species in the Polyergus lucidus complex from eastern North America, rather than that of the Polyergus breviceps complex from western North America and the European species P. rufescens. These results are discussed in the context of speciation within the genus.

Determination of 3,5-Dinitrosalicylic Acid Hydrazide in Honey by Solid-Phase Extraction-Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry.

ABSTRACT: 3,5-Dinitrosalicylic acid hydrazide (DNSAH) is the metabolite of the antibacterial nitrofuran nifursol. A simple and accurate analytical method to determine DNSAH levels in honey by solid-phase extraction-ultraperformance liquid chromatography-tandem mass spectrometry has been established. The honey sample was hydrolyzed under acidic conditions and derivatized with 2-nitrobenzaldehyde in the dark for 16 h, followed by solid-phase extraction and column chromatography. Detection was performed with an electrospray ionization source and multiple reaction monitoring mode and was quantified by the internal standard method. The detection and quantitative limits of DNSAH in honey were 0.1 and 0.3 µg/kg, respectively. Good linear relationships between peak areas and mass concentrations of the analyte were achieved in the range of approximately 0.1 to 200 µg/L, with the correlation coefficient >0.9991. Under the concentration levels of 0.2, 0.5, 1.0, and 2.0 µg/kg, the average recovery rate were between 98.5 and 102.3%, and the relative standard deviations were between 1.1 and 5.4%. The results indicate that the method is simple, rapid, sensitive, and reproducible and can be used to determine DNSAH levels in different types of honey.

Air-assisted liquid-liquid microextraction based on solidification of floating deep eutectic solvent for the analysis of ultraviolet filters in water samples by high performance liquid chromatography with the aid of response surface methodology.

For this work, a novel air-assisted liquid-liquid microextraction based on solidification of floating deep eutectic solvent (AA-LLME-SFDES), coupled with a high performance liquid chromatography (HPLC) method was developed for the detection of benzophenone and salicylate ultraviolet filters in water samples. Three types of fatty acid-based hydrophobic deep eutectic solvents (DESs) with low viscosity, low-density, and melting point close to room temperature were prepared and employed as extraction solvents. This air-assisted liquid-liquid microextraction was carried out in a glass centrifuge tube. Subsequently, the glass tube was introduced into ice-water bath and held for 3 min, during which the upper DES phase was solidified. The water phase was easily extracted using a syringe equipped with a long needle, and later, the glass tube was removed from ice-water bath. The solidified DES phase was immediately melted at room temperature and used for HPLC analysis. The response surface methodology was employed to optimize some influencing parameters such as the volume of the extraction solvent, the pH value of sample solution, the number of extraction cycles, and the addition of salt. A quadratic model, namely a central composite design, was used to replace the conventional single factor analysis. It was found that under optimal conditions, the limits of determination and quantification were 0.045-0.54 µg L-1 and 0.15-2.0 µg L-1, respectively. The relative standard deviations for inter-day (n = 5) and intra-day (n = 5) precision were ≤ 4.2%, whereas the enrichment factors for the ultraviolet filters were obtained from 41 to 50. Furthermore, this novel method was successfully employed for the detection of benzophenone and salicylate ultraviolet filters from real water samples. The recoveries ranged from 87.5% to 105.8%, whereas the RSDs were lower than 3.6%.

Ion-selective electrode integrated in small-scale bioreactor for continuous intracellular pH determination in Lactobacillus plantarum.

The aim of the present study was to develop an ion-selective electrode method for the continuous determination of the intracellular pH in Lactobacillus plantarum using a small-scale bioreactor. This method employed a salicylate-selective electrode basing on the distribution of salicylic acid across the cytoplasmic membrane. This developed electrode responded to salicylate concentrations above 20 µmol/L with a Nernstian sensitivity. The energized and concentrated cells were added into a thermostated small-scale bioreactor that contained the salicylate anions dissolved in a 100 mmol/L potassium phosphate buffer at different pH values. The changes in salicylate concentration that occurred in the medium containing bacterial suspension were measured as a voltage change. The cells of Lactobacillus plantarum showed maintenance of pH homeostasis at the studied pH ranging from 4.0 to 7.0, and they kept a neutral intracellular pH up to 5.8. The simplicity of the measuring preparation and the relatively low cellular concentration, as well as the advantages of the small-scale bioreactor, lead us to believe that the described method can facilitate the study of the physicochemical factors on the intracellular pH of lactic acid bacteria using a single pH probe in one method.

Structure and Chemical Analysis of Major Specialized Metabolites Produced by the Lichen Evernia prunastri.

We performed comparative profiling of four specialized metabolites in the lichen Evernia prunastri, collected at three different geographic locations, California and Maine, USA, and Yoshkar Ola, Mari El, Russia. Among the compounds produced at high concentrations that were identified in all three specimens, evernic acid, usnic acid, lecanoric acid and chloroatranorin, evernic acid was the most abundant. Two depsidones, salazinic acid and physodic acid, were detected in the Yoshkar-Ola collection only. The crystalline structure of evernic acid (2-hydroxy-4-[(2-hydroxy-4-methoxy-6-methylbenzoyl)oxy]-6-methylbenzoate) (hmb) revealed two crystallographically and conformationally distinct hmb anions, along with two monovalent sodium atoms. One hmb moiety contained an exotetradentate binding mode to sodium, whereas the other exhibited an exohexadentate binding mode to sodium. Embedded edge-sharing {Na2 O8 }n sodium-oxygen chains connected the hmb anions into the full three-dimensional crystal structure of the title compound. The crystal used for single-crystal X-ray diffraction exhibited non-merohedral twinning. The data suggest the importance of the acetyl-polymalonyl pathway products to processes of maintaining integrity of the lichen holobiont community.