Gene expression analysis of drought tolerance and cuticular wax biosynthesis in diploid and tetraploid induced wallflowers.

Whole-genome doubling leads to cell reprogramming, upregulation of stress genes, and establishment of new pathways of drought stress responses in plants. This study investigated the molecular mechanisms of drought tolerance and cuticular wax characteristics in diploid and tetraploid-induced Erysimum cheiri. According to real-time PCR analysis, tetraploid induced wallflowers exhibited increased expression of several genes encoding transcription factors (TFs), including AREB1 and AREB3; the stress response genes RD29A and ERD1 under drought stress conditions. Furthermore, two cuticular wax biosynthetic pathway genes, CER1 and SHN1, were upregulated in tetraploid plants under drought conditions. Leaf morphological studies revealed that tetraploid leaves were covered with unique cuticular wax crystalloids, which produced a white fluffy appearance, while the diploid leaves were green and smooth. The greater content of epicuticular wax in tetraploid leaves than in diploid leaves can explain the decrease in cuticle permeability as well as the decrease in water loss and improvement in drought tolerance in wallflowers. GC‒MS analysis revealed that the wax components included alkanes, alcohols, aldehydes, and fatty acids. The most abundant wax compound in this plant was alkanes (50%), the most predominant of which was C29. The relative abundance of these compounds increased significantly in tetraploid plants under drought stress conditions. These findings revealed that tetraploid-induced wallflowers presented upregulation of multiple drought-related and wax biosynthesis genes; therefore, polyploidization has proved useful for improving plant drought tolerance.

A long non-coding RNA functions as a competitive endogenous RNA to modulate TaNAC018 by acting as a decoy for tae-miR6206.

Increasing evidence indicates a strong correlation between the deposition of cuticular waxes and drought tolerance. However, the precise regulatory mechanism remains elusive. Here, we conducted a comprehensive transcriptome analysis of two wheat (Triticum aestivum) near-isogenic lines, the glaucous line G-JM38 rich in cuticular waxes and the non-glaucous line NG-JM31. We identified 85,143 protein-coding mRNAs, 4,485 lncRNAs, and 1,130 miRNAs. Using the lncRNA-miRNA-mRNA network and endogenous target mimic (eTM) prediction, we discovered that lncRNA35557 acted as an eTM for the miRNA tae-miR6206, effectively preventing tae-miR6206 from cleaving the NAC transcription factor gene TaNAC018. This lncRNA-miRNA interaction led to higher transcript abundance for TaNAC018 and enhanced drought-stress tolerance. Additionally, treatment with mannitol and abscisic acid (ABA) each influenced the levels of tae-miR6206, lncRNA35557, and TaNAC018 transcript. The ectopic expression of TaNAC018 in Arabidopsis also improved tolerance toward mannitol and ABA treatment, whereas knocking down TaNAC018 transcript levels via virus-induced gene silencing in wheat rendered seedlings more sensitive to mannitol stress. Our results indicate that lncRNA35557 functions as a competing endogenous RNA to modulate TaNAC018 expression by acting as a decoy target for tae-miR6206 in glaucous wheat, suggesting that non-coding RNA has important roles in the regulatory mechanisms responsible for wheat stress tolerance.

Changes in the structure and enzyme binding of starches during in vitro enzymatic hydrolysis using mammalian mucosal enzyme mixtures.

Starches are hydrolyzed into monosaccharides by mucosal α-glucosidases in the human small intestine. However, there are few studies assessing the direct digestion of starch by these enzymes. The objective of this study was to investigate the changes in the structure and enzyme binding of starches during in vitro hydrolysis by mammalian mucosal enzymes. Waxy maize (WMS), normal maize (NMS), high-amylose maize (HAMS), waxy potato (WPS), and normal potato (NPS) starches were examined. The order of the digestion rate was different compared with other studies using a mixture of pancreatic α-amylase and amyloglucosidase. NPS was digested more than other starches. WPS was more digestible than WMS. Hydrolyzed starch from NPS, NMS, WPS, WMS, and HAMS after 24 h was 66.4, 64.2, 61.7, 58.7, and 46.2 %, respectively. Notably, a significant change in the morphology, reduced crystallinity, and a decrease in the melting enthalpy of the three starches (NPS, NMS, and WPS) after 24 h of hydrolysis were confirmed by microscopy, X-ray diffraction, and differential scanning calorimetry, respectively. The bound enzyme fraction of NPS, NMS, and WPS increased as hydrolysis progressed. In contrast, HAMS was most resistant to hydrolysis by mucosal α-glucosidases in terms of digestibility, changes in morphology, crystallinity, and thermal properties.

Investigation of the influence of minor components and fatty acid profile of oil on properties of beeswax and stearic acid-based oleogels.

Understanding the impact of minor components and the fatty acid profile of oil on oleogel properties is essential for optimizing their characteristics. Considering the scarcity of literature addressing this aspect, this study aimed to explore the correlation between these factors and the properties of beeswax and stearic acid-based oleogels derived from rice bran oil and sesame oil. Minor oil components were modified by stripping the oil, heating the oil with water, and adding ß-sitosterol. Oleogels were then prepared using a mixture of beeswax and stearic acid (3:1, w/w) at a concentration of 11.74 % (w/w). The properties of oils and oleogels were evaluated. The findings indicated that minor components and fatty acid composition of the oils substantially influence the oleogel properties. Removing minor components by stripping resulted in smaller and less uniformly distributed crystals and less oil binding capacity compared to the oleogels prepared from untreated oils. A moderate amount of minor components exhibited a significant influence on oleogel properties. The addition of ß-sitosterol did not show any influence on oleogel properties except for the oleogel made from untreated oil blend added with ß-sitosterol which had more uniform crystals in the microstructure and demonstrated better rheological stability when stored at 5 °C for two months. The oil composition did not show any influence on the thermal and molecular properties of oleogels. Consequently, the oleogel formulation derived from the untreated oil blend enriched with ß-sitosterol was identified as the optimal formula for subsequent development. The findings of this study suggest that the physical and mechanical properties as well as the oxidative stability of beeswax and stearic acid-based oleogels are significantly affected by the minor constituents and fatty acid composition of the oil. Moreover, it demonstrates that the properties of oleogels can be tailored by modifying oil composition by blending different oils.

Another Brick in the Wall of Tear Film Insights Added Through the Total Synthesis and Biophysical Profiling of anteiso-Branched Wax and Cholesteryl Esters.

The tear film lipid layer (TFLL) plays a vital part in maintenance of ocular health and represents a unique biological barrier comprising unusual and specialized lipid classes and species. The wax and cholesteryl esters (WEs and CEs) constitute roughly 80-90% of the TFLL. The majority of species in these lipid classes are branched and it is therefore surprising that the synthesis and properties of the second largest category of species, i.e., the anteiso-branched species, remain poorly characterized. In this study, we have developed a total synthesis route and completed a detailed NMR spectroscopic characterization of two common anteiso-branched species, namely: (22S)-22-methyltetracosanyl oleate and cholesteryl (22'S)-22'-methyltetracosanoate. In addition, we have studied their structural properties in the bulk state by wide-angle and small-angle X-ray scattering and their behavior at the aqueous interface using Langmuir monolayer techniques. A comparison to the properties displayed by iso-branched and straight-chain analogues indicate that branching patterns lead to distinct properties in the CE and WE lipid classes. Overall, this study complements the previous work in the field and adds another important brick in the tear film insights wall.

Sustainable beeswax modified cellulose paper for the determination of tricyclic antidepressants in biofluids.

This article describes the synthesis of sorptive phases for bioanalysis based on the modification of cellulose paper with natural beeswax as sorbent, resulting in a substrate completely renewable and sustainable. The preparation of the sorptive phases consisted of the dissolution of beeswax in hexane, followed by its drop-casting on cellulose paper and subsequent evaporation of the solvent. The beeswax modification of paper renders it hydrophobic, enabling the extraction of the target analytes, i.e., imipramine, desipramine, amitriptyline and trimipramine, via hydrophobic interactions. The main variables affecting the extraction performance were investigated (e.g., pH, ionic strength, extraction time, eluent composition, agitation speed). The analytical workflow combines a straightforward sampling, simultaneous extraction of 30 samples in 1 h, and the rapid (<2 min) determination of the analytes via direct infusion mass spectrometry. The method provided limits of detection in the range 2.0 and 3.2 µg L-1, and the precision, expressed as relative standard deviation, was better than 5.4 % and 8.5 % for intra and inter-day analyses, respectively. The accuracy, in terms of relative recovery, ranged from 90 % to 121 % using saliva as model biofluid.

Morphology and chemical composition of Taiwan oil millet (Eccoilopus formosanus) epicuticular wax.

MAIN CONCLUSION: Taiwan oil millet has two types of epicuticular wax: platelet wax composed primarily of octacosanol and filament wax constituted essentially by the singular compound of octacosanoic acid. Taiwan oil millet (TOM-Eccoilopus formosanus) is an orphan crop cultivated by the Taiwan indigenous people. It has conspicuous white powder covering its leaf sheath indicating abundant epicuticular waxes, that may contribute to its resilience. Here, we characterized the epicuticular wax secretion in TOM leaf blade and leaf sheath using various microscopy techniques, as well as gas chromatography to determine its composition. Two kinds of waxes, platelet and filaments, were secreted in both the leaf blades and sheaths. The platelet wax is secreted ubiquitously by epidermal cells, whereas the filament wax is secreted by a specific cell called epidermal cork cells. The newly developed filament waxes were markedly re-synthesized by the epidermal cork cells through papillae protrusions on the external periclinal cell wall. Ultrastructural images of cork cell revealed the presence of cortical endoplasmic reticulum (ER) tubules along the periphery of plasma membrane (PM) and ER-PM contact sites (EPCS). The predominant wax component was a C28 primary alcohol in leaf blade, and a C28 free fatty acid in the leaf sheath, pseudopetiole and midrib. The wax morphology present in distinct plant organs corresponds to the specific chemical composition: platelet wax composed of alcohols exists mainly in the leaf blade, whereas filament wax constituted mainly by the singular compound C28 free fatty acids is present abundantly in leaf sheath. Our study clarifies the filament wax composition in relation to a previous study in sorghum. Both platelet and filament waxes comprise a protection barrier for TOM.

Feasibility of replacing waxy rice with waxy or sweet-waxy corn viewed from the structure and physicochemical properties of starches.

To explore the feasibility of substituting waxy rice with waxy or sweet-waxy corn, eight varieties of waxy and sweet-waxy corns were selected, including three self-cultivated varieties (Feng nuo 168, Feng nuo 211, and Feng nuo 10). Their starches were isolated and used as research objects, and commercially available waxy rice starch (CAWR) and waxy corn starch (CAWC) were used as controls. X-ray diffraction, scanning electron microscopy, differential scanning calorimetry, rapid viscosity analyzer, and rotational rheometer were used to analyze their physicochemical and structural characteristics. The morphologies of all corn starch granules were generally oval or round, with significant differences in particle size distributions. All ten starches exhibited a typical A-type crystal structure; however, their relative crystallinity varied from 20.08% to 31.43%. Chain length distribution analysis showed that the A/B ratio of Jing cai tian nuo 18 and Feng nuo 168 was similar to that of CAWR. Peak viscosities of corn starches were higher than that of CAWR, except for Feng nuo 10, while their setback values were lower than that of CAWR. Except for Feng nuo 10, the paste transparency of corn starches was higher than that of CAWR (10.77%), especially for Jing cai tian nuo 18 (up to 24%). In summary, Jing cai tian nuo 18 and Feng nuo 168 are promising candidates to replace CAWR in developing various rice-based products.

Paraffin waxes in the North-Western Mediterranean Sea: A comprehensive assessment in the Pelagos Sanctuary, a Specially Protected Area of Mediterranean Importance.

Paraffin waxes are widely recognized as emerging marine pollutants, even their classification by the recent monitoring programs and the knowledge of their occurrence, and sources of contamination in marine ecosystems are poorly defined and reported. Wax presence and distribution have been evaluated in different environmental compartments in the Pelagos Sanctuary (Mediterranean Sea) floating on the sea surface and stranded on beaches, focussing on their characterization, accumulation areas and pollution inputs. More than 2500 yellow paraffin residues were detected and analysed in the study area showing a prevailing dimension smaller than 5 mm. The Genoa Canyon and the waters facing Gorgona Island resulted in the more polluted areas representing two distinct hotspots of wax accumulation potentially related to the high density of tanker vessels sailing to and from the harbour of Genova and Livorno. Higher concentrations of beached particles were found along the Tuscan coast (11 items/100 m) and on Pianosa Island (110 items/m2). This study gives valuable insights into paraffin wax pollution in the Pelagos Sanctuary, emphasizing the need for harmonized monitoring and detection methods to elucidate the potential impacts on marine organisms. Moreover, mitigating actions are crucial to prevent and curb the waxes pollution of marine ecosystems.

Self-assembled, disordered structural color from fruit wax bloom.

Many visually guided frugivores have eyes highly adapted for blue sensitivity, which makes it perhaps surprising that blue pigmented fruits are not more common. However, some fruits are blue even though they do not contain blue pigments. We investigate dark pigmented fruits with wax blooms, like blueberries, plums, and juniper cones, and find that a structural color mechanism is responsible for their appearance. The chromatic blue-ultraviolet reflectance arises from the interaction of the randomly arranged nonspherical scatterers with light. We reproduce the structural color in the laboratory by recrystallizing wax bloom, allowing it to self-assemble to produce the blue appearance. We demonstrate that blue fruits and structurally colored fruits are not constrained to those with blue subcuticular structure or pigment. Further, convergent optical properties appear across a wide phylogenetic range despite diverse morphologies. Epicuticular waxes are elements of the future bioengineering toolbox as sustainable and biocompatible, self-assembling, self-cleaning, and self-repairing optical biomaterials.

Reduction Foodborne Pathogens and Surrogate Microorganism on Citrus Fruits after Lab- and Pilot-scale Finishing Wax Application.

After finishing waxes are applied, citrus fruits are typically dried at 32-60°C for 2-3 min before final packing. The survival of Listeria monocytogenes, Salmonella, and Enterococcus faecium NRRL B-2354 was evaluated under laboratory conditions on lemons after applying one of four finishing waxes (F4, F6, F8, and F15) followed by an ambient hold or heated (50 or 60°C) drying step. The reduction of inoculated microorganisms during drying was significantly influenced by wax type and temperature, with greater reductions at higher temperatures. Greater reductions after waxing and drying at 60°C were observed with L. monocytogenes (2.84-4.44 log) than with Salmonella (1.65-3.67 log), and with Salmonella than with E. faecium (0.99-2.93 log). The survival of Salmonella inoculated at 5.8-5.9 log/fruit on lemons and oranges after applying wax F6 and drying at 60°C was evaluated during storage at 4 and 22°C. The reductions of Salmonella after waxing and drying were 1.7 log; additional reductions during storage at 4 or 22°C were 1.40-1.43 or 0.18-0.29 log, respectively, on waxed lemons, and 0.56-1.02 or 0.54-0.57 log, respectively, on waxed oranges. Under pilot-scale packinghouse conditions with wax F4, mean and minimum reductions of E. faecium ranged from 2.15 to 2.89 and 1.64 to 2.12 log, respectively. However, E. faecium was recovered by whole-fruit enrichment (limit of detection: 0.60 log CFU/lemon) but not by plating (LOD: 1.3 log CFU/lemon) from uninoculated lemons run with or after the inoculated lemons. The findings should provide useful information to establish and implement packinghouse food safety plans.

Analysis of comb-gnawing behavior in Apis cerana cerana (Hymenoptera: Apidae).

Apis cerana cerana exhibits a prominent biological trait known as comb gnawing. In this study, gnawed combs from colonies during different seasons were collected, investigating the comb age and locations of gnawing. Patterns of comb gnawing were recorded, and the effects of 2 factors, namely, comb type and season, on the mass of wax residues and the gnawed surface area were measured. The results revealed that A. c. cerana predominantly gnaws combs that have been used for over 6 months, with gnawing concentrated in the brood-rearing area. In the first 3 seasons, significantly higher masses of wax residues and larger gnawed surface areas were found in greater wax moth larvae (GWML)-infested combs compared to newly built and old combs. Also, there were significantly higher masses and areas gnawed by A. c. cerana in old combs compared to newly built combs in all 4 seasons. Compared to other seasons, it exhibited significantly higher masses and areas resulting from comb-gnawing in newly built or old combs in winter. However, there were no significant differences in the masses of wax residues and surface areas gnawed in GWML-infested combs across the first 3 seasons. In conclusion, this study documented the impact of comb type and season on the comb-gnawing behavior of A. c. cerana, contributing to beekeeping management practices and the current understanding of bee biology.

Carnauba wax-based edible coatings retain quality enhancement of orange (Citrus sinensis cv. Moro) fruits during storage.

Fruit coatings serve a dual purpose in preserving the quality of fruits. Not only do they act as a barrier against water evaporation and fungal infiltration, but they also enhance the fruit's visual appeal in the market. Yet, their influence on the fruit's quality components, which play a crucial role in determining its nutritional value, taste, and overall flavor, has remained relatively unexplored. This study aimed to evaluate the effects of carnauba wax coating on the quality of Moro oranges during storage. The selected fruits were meticulously chosen for uniformity in size. The experiment involved applying carnauba wax, a commonly used type among local producers, at four different concentrations: 0%, 0.5%, 1%, and 1.5%. These treatments were applied during various storage periods, including immediately after fruits were harvested and after 40 and 80 days. Following the application of these treatments, the oranges were stored in a controlled environment (morgue) at a temperature of 4 ± 1 °C. Subsequently, several physicochemical parameters of both the fruit flesh and skin were examined. The results unveiled a decline in the overall ascorbic acid content of the fruits. In terms of phenol content, a general decreasing trend was observed after harvesting. At each sampling interval during storage, the phenol content in uncoated fruits consistently exceeded that of their waxed counterparts. Significant reduction in fruit weight was observed throughout the storage period. Both vitamin C and total acidity levels in the fruit exhibited decreases during the storage period. As time passed, fruit firmness gradually declined, while fruit decay increased during the 40- and 80-day storage periods for untreated Moro oranges. The anthocyanin content showed an increasing trend. The study also unveiled a decline in the antioxidant capacity of citrus fruits during storage. Strong significant positive correlations were observed between total phenol content and key parameters, such as antioxidant activity (0.941**), MDA (0.364*), vitamin C content, and total carbohydrate content (0.475**). Skin radiance showed a perfect correlation with chroma and hue (1.000**). Principal component analysis revealed that the first principal component accounted for 34.27% of the total variance, out of a total of five principal components that explained 77.14% of the variance. Through cluster analysis, the variables were categorized into three distinct groups; one associated with weight loss and another with ion leakage. Considering these findings, carnauba wax-based coating emerges as a promising solution for preserving Moro oranges. It effectively mitigates fruit weight loss and helps maintain fruit firmness during storage, making it a valuable tool for fruit preservation.

Respiratory Exposure to Highly Fluorinated Chemicals via Application of Ski Wax and Related Health Effects.

PURPOSE: Waxes containing per- and polyfluoroalkyl substances (PFAS) are applied to the base of skis and snowboards ("skis") to reduce friction with the snow surface and improve glide. PFAS exposure can adversely impact cardiometabolic, thyroid, liver, kidney, reproductive, and immune health and are associated with increased risk of certain cancers. In the present review, we summarize the state of the science on PFAS exposure from fluorinated ski wax use, including acute respiratory health effects and PFAS concentrations in biological and environmental media collected from ski waxing settings. RECENT FINDINGS: Perfluoroalkyl carboxylic acid (PFCA) concentrations in serum and air collected from professional wax technicians and the rooms where waxes are applied are among the highest of any occupation investigated to date, including the fluorochemical industry. High airborne concentrations of fluorotelomer alcohols contribute to high body burdens of certain PFCAs among ski waxers. Fluorinated ski waxes are a significant source of PFAS exposure for people waxing skis and/or spending time in areas where waxing occurs. We highlight recommendations for future research, policy, and technologies needed to address PFAS exposures from fluorinated wax use.

Chemical compositions and cryo-adhesive probing of the epicuticular wax crystals on fruit surface of wax gourd (Benincasa hispida).

Surface wax crystals play important roles in protecting plants from pest and disease invasions, and UV irradiation. The wax crystals are less probed individually from the fruit surfaces. Herein the morphologies, chemicals and an efficient method to sample the wax blooms of white wax gourd were addressed. Various crystalloids such as rodlets, platelets, fragments, and granules were observed, which stacked as fine wax film covering on wax gourd fruit surface. The wax blooms were effectively removed by cryo-adhesive after consecutive manipulating set by a high-end device with cylinders. Wax crystals were dominated by triterpenols and triterpenol acetates over 61 % of total crystals, followed by vey-long-chain aliphatics. Accordingly, the high-end device with cryo-adhesive provides an efficient approach to selectively probe the wax crystals from those fruits covering wax blooms. The elucidation of morphologies and chemical compositions of wax crystals may help to better understand their regulations on fruit quality traits.

Determination of bisphenols in beeswax based on sugaring out-assisted liquid-liquid extraction: Method development and application in survey, recycling and degradation studies.

The methodology of sugaring out-assisted liquid-liquid extraction (SULLE) coupled with high-performance liquid chromatography-fluorescence detection was devised for quantifying bisphenol A (BPA) and bisphenol B (BPB) in beeswax. The effectiveness of SULLE was methodically explored and proved superior to the salting out-assisted liquid-liquid extraction approach for beeswax sample preparation. The analytical performance underwent comprehensive validation, revealing detection limits of 10 µg/kg for BPA and 20 µg/kg for BPB. The method developed was employed to analyse commercial beeswax (n = 15), beeswax foundation (n = 15) and wild-build comb wax (n = 26) samples. The analysis revealed BPA presence in four commercial beeswax samples and three beeswax foundation samples, with the highest detected residue content being 88 ± 7 µg/kg. For BPB, two beeswax foundation samples were positive, with concentrations below the limits of quantification and 85 ± 4 µg/kg, respectively. No bisphenols were detected in wild-build comb wax. Furthermore, the bisphenol removal efficacy of two recycling methods-boiling in water and methanol extraction-was assessed. The findings indicated that after four recycling cycles using water boiling, 9.6% of BPA and 29.2% of BPB remained in the beeswax. Whereas methanol extraction resulted in approximately 7% residual after one recycling process. A long-term study over 210 days revealed the slow degradation of bisphenols in comb beeswax. This degradation fitted well with a first-order model, indicating half-lives (DT50) of 139 days for BPA and 151 days for BPB, respectively. This research provides the first report on bisphenol contamination in beeswax. The low removal rate during the recycling process and the gradual degradation in beeswax underscore the significance of bisphenol contamination and migration in bee hives along with their potential risk to pollinators warranting concern. Furthermore, the developed SULLE method shows promise in preparing beeswax samples to analyse other analytes.