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The plant cuticle is a complex extracellular lipid barrier that has multiple protective functions. This study investigated cuticle deposition by integrating metabolomics and transcriptomics data gathered from six different maize seedling organs of four genotypes, the inbred lines B73 and Mo17, and their reciprocal hybrids. These datasets captured the developmental transition of the seedling from heterotrophic skotomorphogenic growth to autotrophic photomorphogenic growth, a transition that is highly vulnerable to environmental stresses. Statistical interrogation of these data revealed that the predominant determinant of cuticle composition is seedling organ type, whereas the seedling genotype has a smaller effect on this phenotype. Gene-to-metabolite associations assessed by integrated statistical analyses identified three gene networks associated with the deposition of different elements of the cuticle: cuticular waxes; monomers of lipidized cell wall biopolymers, including cutin and suberin; and both of these elements. These gene networks reveal three metabolic programs that appear to support cuticle deposition, including processes of chloroplast biogenesis, lipid metabolism, and molecular regulation (e.g. transcription factors, post-translational regulators, and phytohormones). This study demonstrates the wider physiological metabolic context that can determine cuticle deposition and lays the groundwork for new targets for modulating the properties of this protective barrier. [ABSTRACT FROM AUTHOR]

The maize glossy2 and glossy2-like genes are homologs, which encode proteins that belong to the BAHD family of acyltransferases. In planta genetic studies have demonstrated that these genes may be involved in the elongation of very long chain fatty acids (VLCFAs) that are precursors of the cuticular wax fraction of the plant cuticle. VLCFAs are synthesized by a fatty acyl-CoA elongase complex (FAE) that consists of four component enzymes. Previously, we functionally identified the maize FAE component enzymes by their ability to complement haploid Saccharomyces cerevisiae strains that carry lethal deletion alleles for each FAE component enzyme. In this study we used these complemented haploid strains and wild-type diploid strains to evaluate whether the co-expression of either GLOSSY2 or GLOSSY2-LIKE with individualmaize FAE component enzymes affects the VLCFA product-profile of the FAE system. Wild-type diploid strains produced VLCFAs of up to 28-carbon chain length. Co-expression of GLOSSY2 orGLOSSY2-LIKE with a combination of maize 3-ketoacyl-CoA synthases stimulated the synthesis of longer VLCFAs, up to 30-carbon chain lengths. However, such results could not be recapitulated when these co-expression experiments were conducted in the yeast haploid mutant strains that lacked individual components of the endogenous FAE system. Specifically, lethal yeast mutant strains that are genetically complemented by the expression of maize FAE-component enzymes produce VLCFAs that range between 20- and 26-carbon chain lengths. However, expressing either GLOSSY2 or GLOSSY2-LIKE in these complemented strains does not enable the synthesis of longer chain VLCFAs. These results indicate that the apparent stimulatory role of GLOSSY2 or GLOSSY2-LIKE to enable the synthesis of longer chain VLCFAs in diploid yeast cells may be associated with mixing plant enzyme components with the endogenous FAE complex. [ABSTRACT FROM AUTHOR]

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. Key message: LncRNA35557 functions as a competing endogenous RNA to modulate TaNAC018 expression by acting as a decoy target for tae-miR6206 in glaucous wheat. [ABSTRACT FROM AUTHOR]

Gas chromatography coupled to mass spectrometry (GC-MS) has been widely used, and along with other methodologies such as derivatization, is a powerful tool to analyze phytocompounds, including cuticular wax components. The loss of structural information upon fragmentation of waxes makes it difficult to interpret and identify the GC-MS mass spectra of these compounds with the traditional methods, as digital libraries. Therefore, as a new complementary alternative to traditional methods of identifying aliphatic acyclic lipids based on mass spectra, we present the new software WaxAlly in this technical note. WaxAlly is a new software created based on simple algorithms, enabling the rapid recognition by the user of eight aliphatic acyclic lipid classes, including alkanes, alkenes, aldehydes, ketones, esters, and TMS derivatives of free fatty acids, primary and secondary alcohols, and their homologues with carbon chains varying between 10 and 100 carbons. Additionally, the WaxAlly software provides a section for data organization, internet comparison to NIST and PubChem databases, and academic information about mass fragmentation of acyclic lipids. The software has proven to be a very useful complementary technique in identifying plant wax lipid homologues, and new fragmentation patterns of lipid classes can be added in the future to improve the program.

Cuticular waxes coating leaf surfaces can help plants tolerate drought events by reducing non-stomatal water loss. Despite their role in drought tolerance, little is known about how cuticular wax composition has changed during breeding in Canadian bread wheat (Triticum aestivum L.) varieties. To fill in this gap, flag leaves of the Canadian Heritage Bread Wheat Panel, which include 30 varieties released between 1842 and 2018, were surveyed to determine if and how cuticular wax composition in wheat has changed at two breeding ecozones over this period. Following this, a subset of varieties was subjected to drought conditions to compare their responses. As expected, modern varieties outperformed old varieties with a significantly larger head length and reaching maturity earlier. Yet, when challenged with drought, old varieties were able to significantly increase the accumulation of β-diketones to a higher extent than modern varieties. Furthermore, RNAseq was performed on the flag leaf of four modern varieties to identify potential markers that could be used for selection of higher accumulation of cuticular waxes. This analysis revealed that the W1 locus is a good candidate for selecting higher accumulation of β-diketones. These findings indicate that the variation in cuticular waxes upon drought could be further incorporated in breeding of future bread wheat varieties.

Both leaves and petals are covered in a cuticle, which itself contains and is covered by cuticular waxes. The waxes perform various roles in plants’ lives, and the cuticular composition of leaves has received much attention. To date, the cuticular composition of petals has been largely ignored. Being the outermost boundary between the plant and the environment, the cuticle is the first point of contact between a flower and a pollinator, yet we know little about how plant–pollinator interactions shape its chemical composition. Here, we investigate the general structure and composition of floral cuticular waxes by analysing the cuticular composition of leaves and petals of 49 plant species, representing 19 orders and 27 families. We show that the flowers of plants from across the phylogenetic range are nearly devoid of wax crystals and that the total wax load of leaves in 90% of the species is higher than that of petals. The proportion of alkanes is higher, and the chain lengths of the aliphatic compounds are shorter in petals than in leaves. We argue these differences are a result of adaptation to the different roles leaves and petals play in plant biology.

To investigate the influence of heat treatment (HT) on Satsuma mandarin fruit's postharvest quality and cuticle composition, we immersed the fruit for 3 min in hot water at 52°C and subsequently stored them at room temperature (25°C) for 28 days, and fruit quality parameters, such as good fruit rate, weight loss rate, firmness, total soluble solids, total titratable acidity, and ascorbic acid content, were monitored. Additionally, changes in the peel's cuticle composition were analyzed, and wax crystal morphologies on the fruit surface were examined using scanning electron microscopy (SEM). The findings revealed that appropriate HT effectively preserved fruit quality. The main compositions of wax and cutin on the fruit's surface remained consistent between the HT and the CK during storage. The total content of wax and cutin initially increased, peaking on the 14th day of storage, and then decreased, falling below the levels observed on day 0. Notably, the total amount of cutin in the HT group exceeded that of the control group. Specifically, ω‐hydroxy fatty acids with mid‐chain oxo groups and mid‐oh‐ω‐hydroxy fatty acids constituted approximately 90% of the total cutin content. Moreover, the HT group exhibited higher (p < 0.05) total wax content in relation to the control. Fatty acids and alkanes were the predominant components, accounting for approximately 87.5% of the total wax. SEM analysis demonstrated that HT caused wax crystals to melt and redistribute, effectively filling wax gaps. It suggests that HT holds promising potential as a green, safe, and eco‐friendly commercial treatment for preserving the postharvest quality of Satsuma mandarin. Practical Application: In this study, Satsuma citrus (Citrus unshiu) underwent heat treatment (HT) and was subsequently preserved at room temperature (25°C) for 28 days. The findings revealed that HT significantly improved fruit quality compared to the control group. These findings provide valuable insights into the advancement of eco‐friendly and pollution‐free citrus preservation methods, offering essential strategies and process parameters for their practical application. [ABSTRACT FROM AUTHOR]

SUMMARY: With climate change, an aggravation in summer drought is expected in the Mediterranean region. To assess the impact of such a future scenario, we compared the response of Quercus pubescens, a drought‐resistant deciduous oak species, to long‐term amplified drought (AD) (partial rain exclusion in natura for 10 years) and natural drought (ND). We studied leaf physiological and physico‐chemical trait responses to ND and AD over the seasonal cycle, with a focus on chemical traits including major groups of central (photosynthetic pigments and plastoquinones) and specialized (tocochromanols, phenolic compounds, and cuticular waxes) metabolites. Seasonality was the main driver of all leaf traits, including cuticular triterpenoids, which were highly concentrated in summer, suggesting their importance to cope with drought and thermal stress periods. Under AD, trees not only reduced CO2 assimilation (−42%) in summer and leaf concentrations of some phenolic compounds and photosynthetic pigments (carotenoids from the xanthophyll cycle) but also enhanced the levels of other photosynthetic pigments (chlorophylls, lutein, and neoxanthin) and plastochromanol‐8, an antioxidant located in chloroplasts. Overall, the metabolomic adjustments across seasons and drought conditions reinforce the idea that Q. pubescens is highly resistant to drought although significant losses of antioxidant defenses and photoprotection were identified under AD. Significance Statement: Climate change will increase drought episodes, especially in the Mediterranean region. This study investigated Quercus pubescens response to long‐term amplified drought (AD) in natura, focusing on leaf physiological and physico‐chemical traits across seasons and AD. It revealed that seasonal variations influenced leaf metabolism, including leaf cuticular compounds, which are important to cope with drought and thermal stress. Also, AD led to a decrease in net photosynthesis as well as losses in antioxidant defense and photoprotection. [ABSTRACT FROM AUTHOR]

Triterpenoids are a group of secondary plant metabolites, with a remarkable pharmacological potential, occurring in the cuticular waxes of the aerial parts of plants. The aim of this study was to analyze triterpenoid variability in the fruits and leaves of three apple cultivars during the growing season and gain new insights into their health-promoting properties and fate during juice and purée production. The identification and quantification of the compounds of interest were conducted using gas chromatography coupled with mass spectrometry. The waxes of both matrices contained similar analytes; however, their quantitative patterns varied: triterpenic acids prevailed in the fruits, while higher contents of steroids and esterified forms were observed in the leaves. The total triterpenoid content per unit area was stable during the growing season; the percentage of esters increased in the later phases of growth. Antioxidative and anti-inflammatory properties were evaluated with a series of in vitro assays. Triterpenoids were found to be the main anti-inflammatory compounds in the apples, while their impact on antioxidant capacity was minor. The apples were processed on a lab scale to obtain juices and purées. The apple purée and cloudy juice contained only some of the triterpenoids present in the raw fruit, while the clear juices were virtually free of those lipophilic compounds. [ABSTRACT FROM AUTHOR]

Cork oak (Quercus suber L.) has high economic value given by its sustainable production of cork, and ecological importance in the Mediterranean region. The species is well adapted to the dry climate, namely through the sclerophyllous nature of its leaves with a well-developed cuticle, including cutin and cuticular waxes that contribute to protection against drought. Leaves of cork oaks were collected along one annual cycle, starting from the young leaves in May to the one-year-old leaves in March. Leaf cutin content and chemical composition were determined by transesterification subsequently to the determination of cuticular waxes, and leaf features, and were analyzed along the leaf cycle. Cutin is a major component of the cuticle, representing on average 72.4% of the cutin and cuticular waxes. Cutin amounted to 71.0 g/1000 g of dry leaves, without significant seasonal mass proportion variation, while cutin coverage increased from May to December (429.7 μg/cm2 and 575.4 μg/cm2, respectively). In contrast, a clear seasonality was found in cuticular wax mass proportion and coverage (18.4 g/1000 g of dry leaves and 113.5 μg/cm2 in May, and 28.5 g/1000 g and 235.2 μg/cm2 in September). Cutin is a glyceridic polyester composed by long-chain acids, mainly ω-hydroxyacids, followed by fatty acids with a few ω-diacids and alcohols, and by a substantial proportion of aromatics. Cutin composition varied along time with a proportional increase in ω-hydroxyacids (45.8% in May; 50.8% in December), and a significant decrease in aromatics (24.2% in May and 8.5% in March). The cuticle seasonal development in the cork oak contributes to protect the leaves and the trees from the dry summer conditions. [ABSTRACT FROM AUTHOR]

Fruit and leaf cuticular waxes are valuable source materials for the isolation of triterpenoids that can be applied as natural antioxidants and anticancer agents. The present study aimed at the semi-preparative fractionation of triterpenoids from cuticular wax extracts of Vaccinium vitis-idaea L. (lingonberry) leaves and fruits and the evaluation of their cytotoxic potential. Qualitative and quantitative characterization of obtained extracts and triterpenoid fractions was performed using HPLC-PDA method, followed by complementary analysis by GC-MS. For each fraction, cytotoxic activities towards the human colon adenocarcinoma cell line (HT-29), malignant melanoma cell line (IGR39), clear renal carcinoma cell line (CaKi-1), and normal endothelial cells (EC) were determined using MTT assay. Furthermore, the effect of the most promising samples on cancer spheroid growth and viability was examined. This study allowed us to confirm that particular triterpenoid mixtures from lingonberry waxes may possess stronger cytotoxic activities than crude unpurified extracts. Fractions containing triterpenoid acids plus fernenol, complexes of oleanolic:ursolic acids, and erythrodiol:uvaol were found to be the most potent therapeutic candidates in the management of cancer diseases. The specificity of cuticular wax extracts of lingonberry leaves and fruits, leading to different purity and anticancer potential of obtained counterpart fractions, was also enclosed. These findings contribute to the profitable utilization of lingonberry cuticular waxes and provide considerable insights into the anticancer effects of particular triterpenoids and pharmacological interactions. [ABSTRACT FROM AUTHOR]

Cuticular waxes comprise the hydrophobic layer that protects crops against nonstomatal water loss and biotic and abiotic stresses. Expanding on our current knowledge of the genes that are involved in cuticular wax biosynthesis and regulation plays an important role in dissecting the processes of cuticular wax metabolism. In this study, we identified the Cer-GN1 barley (Hordeum vulgare L.) mutant that is generated by ethyl methanesulfonate mutagenesis with a glossy spike phenotype that is controlled by a single recessive nuclear gene. A physiological analysis showed that the total cuticular wax loads of Cer-GN1 were one-third that of the progenitor wild-type (WT), and its water loss rate was significantly accelerated (p < 0.05). In addition, Cer-GN1 was defective in the glume's cuticle according to the toluidine blue dye test, and it was deficient in the tubule-shaped crystals which were observed on the glume surfaces by scanning electron microscopy. Using metabolomics and transcriptomics, we investigated the impacts of cuticular wax composition and waxy regulatory genes on the loss of the glaucous wax in the spikes of Cer-GN1. Among the differential metabolites, we found that 16-hydroxyhexadecanoic acid, which is one of the predominant C16 and C18 fatty acid-derived cutin monomers, was significantly downregulated in Cer-GN1 when it was compared to that of WT. We identified two novel genes that are located on chromosome 4H and are downregulated in Cer-GN1 (HvMSTRG.29184 and HvMSTRG.29185) that encode long-chain fatty acid omega-monooxygenase CYP704B1, which regulates the conversion of C16 palmitic acid to 16-hydroxyhexadecanoic acid. A quantitative real-time PCR revealed that the expression levels of HvMSTRG.29184 and HvMSTRG.29185 were downregulated at 1, 4, 8, 12, and 16 days after the heading stage in Cer-GN1 when it was compared to those of WT. These results suggested that HvMSTRG.29184 and HvMSTRG.29185 have CYP704B1 activity, which could regulate the conversion of C16 palmitic acid to 16-hydroxyhexadecanoic acid in barley. Their downregulation in Cer-GN1 reduced the synthesis of the cuticular wax components and ultimately caused the loss of the glaucous wax in the spikes. It is necessary to verify whether HvMSTRG.29184 and HvMSTRG.29185 truly encode a CYP704B1 that regulates the conversion of C16 palmitic acid to 16-hydroxyhexadecanoic acid in barley. [ABSTRACT FROM AUTHOR]

Fruit extracts of the Chaenomeles species are a rich source of compounds having health-promoting properties, while their distribution between the species and cultivars varies significantly depending on both genotype and environmental threats. This study aimed at discovering antioxidant and antimicrobial potential of the secondary metabolites of fruit and waxes of fruit cuticular of introduced Ch. cathayensis and Ch. × californica plants. The sum of detected polyphenols in the isopropanolic fruit extracts varied slightly between the species, while significant excesses in indices were seen for both species peel extracts as compared to pulp extracts. Antimicrobial assays carried out by disc diffusion method showed notable activity of the fruit peel and pulp extracts of both species against all tested Gram-negative and Gram-positive bacterial strains, and two Candida strains as well. Pseudomonas aeruginosa strain was the most resistant to the action of both fruit extracts, especially peel extracts of Ch. cathayensis fruits. As identified by gas chromatography-mass spectrometry (GC-MS) assays, chloroformic extracts from the fruits of cuticular waxes of Ch. cathayensis and Ch. × californica contained six prevailing fractions: aldehydes, alkanes, alcohols, esters, fatty acids and various terpenoids. The predominant compounds were tetrapentacontane (21.8% of total amount) and heptacosanal (23.1% of total), respectively in the cuticular waxes of Ch. cathayensis and Ch. × californica. Cinnamaldehyde, cis-9-hexadecenal, hexadecanoic acid, oleic acid, olean-12-ene-3,28-diol (3. beta), lupeol, diisooctyl phthalate, 9-octadecenoic acid, 1,2,3-propanetriyl ester, 1,3,12-nonadecatriene-5,14-diol and some other identified compounds are well-known for their bioactivity, indicating the feasibility of studying the antimicrobial potential of plant fruits.

Quercus suber L. (cork oak) leaves were analyzed along one annual cycle for cuticular wax content and chemical composition. This species, well adapted to the long dry summer conditions prevailing in the Mediterranean, has a leaf life span of about one year. The cuticular wax revealed a seasonal variation with a coverage increase from the newly expanded leaves (115.7 µg/cm2 in spring) to a maximum value in fully expanded leaves (235.6 µg/cm2 after summer). Triterpenoids dominated the wax composition throughout the leaf life cycle, corresponding in young leaves to 26 µg/cm2 (22.6% of the total wax) and 116.0 µg/cm2 (49% of the total wax) in mature leaves, with lupeol constituting about 70% of this fraction. The total aliphatic compounds increased from 39 µg/cm2 (young leaves) to 71 µg/cm2 (mature leaves) and then decreased to 22 µg/cm2 and slightly increased during the remaining period. The major aliphatic compounds were fatty acids, mostly with C16 (hexadecanoic acid) and C28 (octacosanoic acid) chain lengths. Since pentacyclic triterpenoids are located almost exclusively within the cutin matrix (intracuticular wax), the increase in the cyclic-to-acyclic component ratio after summer shows an extensive deposition of intracuticular waxes in association with the establishment of mechanical and thermal stability and of water barrier properties in the mature leaf cuticle. [ABSTRACT FROM AUTHOR]

Drought is one of the main abiotic stresses limiting the quality and yield of citrus. Cuticular waxes play an important role in regulating plant drought tolerance and water use efficiency (WUE). However, the contribution of cuticular waxes to drought tolerance, WUE and the underlying molecular mechanism is still largely unknown in citrus. 'Longhuihong' (MT) is a bud mutant of 'Newhall' navel orange with curly and bright leaves. In this study, significant increases in the amounts of total waxes and aliphatic wax compounds, including n-alkanes, n-primary alcohols and n-aldehydes, were overserved in MT leaves, which led to the decrease in cuticular permeability and finally resulted in the improvements in drought tolerance and WUE. Compared to WT leaves, MT leaves possessed much lower contents of malondialdehyde (MDA) and hydrogen peroxide (H2O2), significantly higher levels of proline and soluble sugar, and enhanced superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities under drought stress, which might reduce reactive oxygen species (ROS) damage, improve osmotic regulation and cell membrane stability, and finally, enhance MT tolerance to drought stress. Transcriptome sequencing results showed that seven structural genes were involved in wax biosynthesis and export, MAPK cascade, and ROS scavenging, and seven genes encoding transcription factors might play an important role in promoting cuticular wax accumulation, improving drought tolerance and WUE in MT plants. Our results not only confirmed the important role of cuticular waxes in regulating citrus drought resistance and WUE but also provided various candidate genes for improving citrus drought tolerance and WUE. [ABSTRACT FROM AUTHOR]

The plant cuticle, as a lipid membrane covering aerial plant surfaces, functions primarily against uncontrolled water loss. Herein, the cuticle chemical composition and the transpiration of wampee fruit (Clausena lansium (Lour.) Skeels) at the green, turning, and yellow stages in cultivars of "Jixin" and "Tianhuangpi" were comprehensively studied. The coverage of wax and cutin monomers per unit of fruit surface area at the green stage was lower in "Jixin" than in "Tianhuangpi" and increased gradually during development. Cutin monomers accumulated ranging from 22.5 μg cm−2 (green) to 52.5 μg cm−2 (turning) in "Jixin" and from 36.5 μg cm−2 (green) to 81.7 μg cm−2 (yellow) in "Tianhuangpi." The total composition of waxes ranged between 6.0 μg cm−2 (green) and 11.1 μg cm−2 (turning) in "Jixin," while they increased from 7.4 μg cm−2 (green) to 16.7 μg cm−2 (yellow) in "Tianhuangpi." Cutin monomers were dominated by ω-, mid-dihydroxy fatty acids (over 40%), followed by multiple monomers of α,ω-dicarboxylic acids with or without added groups, α-monocarboxylic acids with or without ω- or mid-chain hydroxy or mid-epoxy groups, primary alcohols, and phenolics. The very-long-chain (VLC) aliphatic pattern of cuticular waxes was prominently composed of n -alkanes (ranging from 21.4% to 39.3% of total wax content), fatty acids, primary alcohols, and aldehydes. The cyclic waxes were dominated by triterpenoids (between 23.9 and 51.2%), sterols, and phenolics. Water loss in wampee fruit exhibited linear changes over time, indicating an overall monofunctional barrier to transpiration. Permeance for water in wampee fruit was higher at the green stage than at the yellow stage in both "Jixin" and "Tianhuangpi," which showed a negative correlation with the changes of VLC n -alkanes. The results showed the cuticular chemicals, including cutin monomers and waxes, in wampee fruit and further indicated the potential contributions of the cuticular chemical composition to the physiological functions in fruits. [ABSTRACT FROM AUTHOR]

Very-long-chain fatty acids (VLCFA) are involved in a number of important plant physiological functions. Disorders in the expression of genes involved in the synthesis of VLCFA lead to a number of phenotypic consequences, ranging from growth retardation to the death of embryos. The elongation of VLCFA in the endoplasmic reticulum (ER) is carried out by multiple elongase complexes with different substrate specificities and adapted to the synthesis of a number of products required for a number of metabolic pathways. The information about the enzymes involved in the synthesis of VLCFA with more than 26 atoms of Carbon is rather poor. Recently, genes encoding enzymes involved in the synthesis of both regular-length fatty acids and VLCFA have been discovered and investigated. Polyunsaturated VLCFA in plants are formed mainly by 20:1 elongation into new monounsaturated acids, which are then imported into chloroplasts, where they are further desaturated. The formation of saturated VLCFA and their further transformation into a number of aliphatic compounds included in cuticular waxes and suberin require the coordinated activity of a large number of different enzymes. [ABSTRACT FROM AUTHOR]

Cuticular waxes can be used in high-value applications, including cosmetics, foods and nutraceuticals, among the others. The extraction process determines their quality and purity that are of particular interest when biocompatibility, biodegradability, flavor and fragrance are the main features required for the final formulations. This study demonstrated that supercritical fluid extraction coupled with fractional separation can represent a suitable alternative to isolate cuticular waxes from vegetable matter that preserve their natural properties and composition, without contamination of organic solvent residues. Operating in this way, cuticular waxes can be considered as a fingerprint of the vegetable matter, where C27, C29 and C31 are the most abundant compounds that characterize the material; the differences are mainly due to their relative proportions and the presence of hydrocarbon compounds possessing other functional groups, such as alcohols, aldehydes or acids. Therefore, selectivity of supercritical fluid extraction towards non-polar or slightly polar compounds opens the way for a possible industrial approach to produce extracts that do not require further purification steps. [ABSTRACT FROM AUTHOR]

The plant cuticle, as a lipid membrane covering aerial plant surfaces, functions primarily against uncontrolled water loss. Herein, the cuticle chemical composition and the transpiration of wampee fruit (Clausena lansium (Lour.) Skeels) at the green, turning, and yellow stages in cultivars of “Jixin” and “Tianhuangpi” were comprehensively studied. The coverage of wax and cutin monomers per unit of fruit surface area at the green stage was lower in “Jixin” than in “Tianhuangpi” and increased gradually during development. Cutin monomers accumulated ranging from 22.5 μg cm−2 (green) to 52.5 μg cm−2 (turning) in “Jixin” and from 36.5 μg cm−2 (green) to 81.7 μg cm−2 (yellow) in “Tianhuangpi.” The total composition of waxes ranged between 6.0 μg cm−2 (green) and 11.1 μg cm−2 (turning) in “Jixin,” while they increased from 7.4 μg cm−2 (green) to 16.7 μg cm−2 (yellow) in “Tianhuangpi.” Cutin monomers were dominated by ω-, mid-dihydroxy fatty acids (over 40%), followed by multiple monomers of α,ω-dicarboxylic acids with or without added groups, α-monocarboxylic acids with or without ω- or mid-chain hydroxy or mid-epoxy groups, primary alcohols, and phenolics. The very-long-chain (VLC) aliphatic pattern of cuticular waxes was prominently composed of n-alkanes (ranging from 21.4% to 39.3% of total wax content), fatty acids, primary alcohols, and aldehydes. The cyclic waxes were dominated by triterpenoids (between 23.9 and 51.2%), sterols, and phenolics. Water loss in wampee fruit exhibited linear changes over time, indicating an overall monofunctional barrier to transpiration. Permeance for water in wampee fruit was higher at the green stage than at the yellow stage in both “Jixin” and “Tianhuangpi,” which showed a negative correlation with the changes of VLC n-alkanes. The results showed the cuticular chemicals, including cutin monomers and waxes, in wampee fruit and further indicated the potential contributions of the cuticular chemical composition to the physiological functions in fruits.

The search for a universal explanation of the altitudinal limit determined by the alpine treeline has given rise to different hypotheses. In this study, we revisited Michaelis’ hypothesis which proposed that an inadequate “ripening” of the cuticle caused a greater transpiration rate during winter in the treeline. However, few studies with different explanations have investigated the role of passive mechanisms of needles for protecting against water loss during winter in conifers at the treeline. To shed light on this, the cuticular transpiration barrier was studied in the transition from subalpine Pinus uncinata forests to alpine tundra at the upper limit of the species in the Pyrenees. This upper limit of P. uncinata was selected here as an example of the ecotones formed by conifers in the temperate mountains of the northern hemisphere. Our study showed that minimum leaf conductance in needles from upper limit specimens was higher than those measured in specimens living in the lower levels of the sub-alpine forest and also displayed lower cuticle thickness values, which should reinforce the seminal hypothesis by Michaelis. Our study showed clear evidence that supports the inadequate development of needle cuticles as one of the factors that lead to increased transpirational water losses during winter and, consequently, a higher risk of suffering frost drought.

The search for a universal explanation of the altitudinal limit determined by the alpine treeline has given rise to different hypotheses. In this study, we revisited Michaelis' hypothesis which proposed that an inadequate "ripening" of the cuticle caused a greater transpiration rate during winter in the treeline. However, few studies with different explanations have investigated the role of passive mechanisms of needles for protecting against water loss during winter in conifers at the treeline. To shed light on this, the cuticular transpiration barrier was studied in the transition from subalpine Pinus uncinata forests to alpine tundra at the upper limit of the species in the Pyrenees. This upper limit of P. uncinata was selected here as an example of the ecotones formed by conifers in the temperate mountains of the northern hemisphere. Our study showed that minimum leaf conductance in needles from upper limit specimens was higher than those measured in specimens living in the lower levels of the sub-alpine forest and also displayed lower cuticle thickness values, which should reinforce the seminal hypothesis by Michaelis. Our study showed clear evidence that supports the inadequate development of needle cuticles as one of the factors that lead to increased transpirational water losses during winter and, consequently, a higher risk of suffering frost drought. [ABSTRACT FROM AUTHOR]

Fruit and leaf cuticular waxes are valuable source materials for the isolation of triterpenoids that can be applied as natural antioxidants and anticancer agents. The present study aimed at the semi-preparative fractionation of triterpenoids from cuticular wax extracts of Vaccinium vitis-idaea L. (lingonberry) leaves and fruits and the evaluation of their cytotoxic potential. Qualitative and quantitative characterization of obtained extracts and triterpenoid fractions was performed using HPLC-PDA method, followed by complementary analysis by GC-MS. For each fraction, cytotoxic activities towards the human colon adenocarcinoma cell line (HT-29), malignant melanoma cell line (IGR39), clear renal carcinoma cell line (CaKi-1), and normal endothelial cells (EC) were determined using MTT assay. Furthermore, the effect of the most promising samples on cancer spheroid growth and viability was examined. This study allowed us to confirm that particular triterpenoid mixtures from lingonberry waxes may possess stronger cytotoxic activities than crude unpurified extracts. Fractions containing triterpenoid acids plus fernenol, complexes of oleanolic:ursolic acids, and erythrodiol:uvaol were found to be the most potent therapeutic candidates in the management of cancer diseases. The specificity of cuticular wax extracts of lingonberry leaves and fruits, leading to different purity and anticancer potential of obtained counterpart fractions, was also enclosed. These findings contribute to the profitable utilization of lingonberry cuticular waxes and provide considerable insights into the anticancer effects of particular triterpenoids and pharmacological interactions.

Fruit extracts of the Chaenomelesspecies are a rich source of compounds having health-promoting properties, while their distribution between the species and cultivars varies significantly depending on both genotype and environmental threats. This study aimed at disco)vering antioxidant and antimicrobial potential of the secondary metabolites of fruit and waxes of fruit cuticular of introduced Ch. cathayensis and Ch. × californica plants. The sum of detected polyphenols in the isopropanolic fruit extracts varied slightly between the species, while significant excesses in indices were seen for both species peel extracts as compared to pulp extracts. Antimicrobial assays carried out by disc diffusion method showed notable activity of the fruit peel and pulp extracts of both species against all tested Gram-negative and Gram-positive bacterial strains, and two Candida strains as well. Pseudomonas aeruginosa strain was the most resistant to the action of both fruit extracts, especially peel extracts of Ch. cathayensisfruits. As identified by gas chromatography-mass spectrometry (GC-MS) assays, chloroformic extracts from the fruits of cuticular waxes of Ch. cathayensis and Ch. × californica contained six prevailing fractions: aldehydes, alkanes, alcohols, esters, fatty acids and various terpenoids. The predominant compounds were tetrapentacontane (21.8% of total amount) and heptacosanal (23.1% of total), respectively in the cuticular waxes of Ch. cathayensis and Ch. × californica. Cinnamaldehyde, cis-9-hexadecenal, hexadecanoic acid, oleic acid, olean-12-ene-3,28-diol (3. beta), lupeol, diisooctyl phthalate, 9-octadecenoic acid, 1,2,3-propanetriyl ester, 1,3,12-nonadecatriene-5,14-diol and some other identified compounds are well-known for their bioactivity, indicating the feasibility of studying the antimicrobial potential of plant fruits. [ABSTRACT FROM AUTHOR]

Valorization of the agri‐food industry by‐products could contribute to curb issues related to food security and environmental problems. Flax and wheat seeds are major products of this industry, but their production is associated with tons of straws that can be valorized for their cuticular and epicuticular waxes. We aimed to determine the organogelation capacity of epicuticular waxes in comparison to cuticular waxes from both flax and wheat straws. Epicuticular waxes from flax and wheat straws have structured canola oil at 2% and 4% (w/w), respectively, whereas cuticular waxes from flax and wheat straws required critical concentrations of 4% and 5% (w/w), respectively. Characterization of the organogelation capacity (onset of crystallization temperature, temperature of phase transition, crystal morphology, solid fat, crystalline structure, and oil binding capacity) was also carried out. The high onset of crystallization temperature (38.1 ± 1.2°C), the phase transition at high temperature (38 ± 1.5°C), and capacity to structure canola oil at low concentration showed that epicuticular wax from flax straw is a promisor fat substitute, presenting organogelation properties comparable to the best results obtained in the literature for other vegetal waxes. [ABSTRACT FROM AUTHOR]

Barrier properties of the hydrophobic plant cuticle depend on its physicochemical composition. The cuticular compounds vary considerably among plant species but also among organs and tissues of the same plant and throughout developmental stages. As yet, these intraspecific modifications at the cuticular wax and cutin level are only rarely examined. Attempting to further elucidate cuticle profiles, we analysed the adaxial and abaxial surfaces of the sclerophyllous leaf and three developmental stages of the drupe fruit of Prunus laurocerasus , an evergreen model plant native to temperate regions. According to gas chromatographic analyses, the cuticular waxes contained primarily pentacyclic triterpenoids dominated by ursolic acid, whereas the cutin biopolyester mainly consisted of 9/10,ω-dihydroxy hexadecanoic acid. Distinct organ- and side-specific patterns were found for cuticular lipid loads, compositions and carbon chain length distributions. Compositional variations led to different structural and functional barrier properties of the plant cuticle, which were investigated further microscopically, infrared spectroscopically and gravimetrically. The minimum water conductance was highlighted at 1 × 10−5 m s−1 for the perennial, hypostomatous P. laurocerasus leaf and at 8 × 10−5 m s−1 for the few-month-living, stomatous fruit suggesting organ-specific cuticular barrier demands. • The cuticle of the evergreen shrub P. laurocerasus displays a high phenotypic plasticity. • Organ-specific patterns of cuticular wax compound classes were detected. • Side-specific shifts were found for carbon chain lengths in leaf cuticular waxes. • Minimum water conductance differed considerably between organs of the same plant. • Temperature-dependent minimum conductance and cuticular permeability were compared. [ABSTRACT FROM AUTHOR]

The cuticular membrane covering almost all aerial plant organs has a primary function in limiting uncontrolled water loss. The guava fruits were collected and this work was done to study the potential contribution of cuticular chemical composition to fruit transpiration after harvest. The detailed cuticular chemical composition, based on gas chromatography together with mass spectrometry, and the transpiration rate determined gravimetrically in guava fruit were characterized in the present study. The predominant wax mixtures were fatty acids and primary alcohols with homologous series of C 16 –C 33 , as well as various pentacyclic triterpenoids with abundant amounts of ursolic acid, maslinic acid and uvaol. The most prominent cutin compounds were C 16 and C 18 ‒type monomers dominated by 9(10),16-diOH-hexadecanoic acid and 9,10-epoxy-ω–OH–octadecanoic acid, respectively. Relatively high water permeability with a value of 5.1 × 10−4 m s−1 was detected for guava fruit. The lower efficiency of the cuticle as barrier to transpiration in guava fruit, as compared to that of other reported fruits, leaves, and petals, was seemingly related to the relatively short average chain-length of acyclic compounds in wax mixtures. These findings provide useful insights linking the chemical composition of the cuticular membrane that covers plant organs to putative physiological roles. • Fatty acids, primary alcohols, and triterpenoids dominated wax mixture of guava fruit. • A mixture of C 16 and C 18 cutin monomers frames the cutin matrix. • Water loss of guava fruit was higher than that of other fruits, leaves and petals. • Low efficient barrier for transpiration in guava fruit due to its cuticular constituents. [ABSTRACT FROM AUTHOR]

The cuticle is composed of cutin and cuticular waxes, and it is the first protective barrier to abiotic and biotic stresses in fruit. In this study, we analysed the composition of and changes in cuticular waxes during fruit development in nectarine (Prunus persica L. Batsch) cultivars, in parallel with their conductance and their susceptibility to Monilinia laxa. The nectarine waxes were composed of triterpenoids, mostly ursolic and oleanolic acids, phytosterols, and very-long-chain aliphatics. In addition, we detected phenolic compounds that were esterified with sugars or with triterpenoids, which are newly described in cuticular waxes. We quantified 42 compounds and found that they changed markedly during fruit development, with an intense accumulation of triterpenoids during initial fruit growth followed by their decrease at the end of endocarp lignification and a final increase in very-long-chain alkanes and hydroxylated triterpenoids until maturity. The surface conductance and susceptibility to Monilinia decreased sharply at the beginning of endocarp lignification, suggesting that triterpenoid deposition could play a major role in regulating fruit permeability and susceptibility to brown rot. Our results provide new insights into the composition of cuticular waxes of nectarines and their changes during fruit development, opening new avenues of research to explore brown rot resistance factors in stone fruit. [ABSTRACT FROM AUTHOR]

Background and Aims The cuticle of a limited number of plant species contains cutan, a chemically highly resistant biopolymer. As yet, the biosynthesis of cutan is not fully understood. Attempting to further unravel the origin of cutan, we analysed the chemical composition of enzymatically isolated cuticular membranes of Agave americana leaves. Methods Cuticular waxes were extracted with organic solvents. Subsequently, the dewaxed cuticular membrane was depolymerized by acid-catalysed transesterification yielding cutin monomers and cutan, a non-hydrolysable, cuticular membrane residue. The cutan matrix was analysed by thermal extraction, flash pyrolysis and thermally assisted hydrolysis and methylation to elucidate the monomeric composition and deduce a putative biosynthetic origin. Key Results According to gas chromatography–mass spectrometry analyses, the cuticular waxes of A. americana contained primarily very-long-chain alkanoic acids and primary alkanols dominated by C32, whereas the cutin biopolyester of A. americana mainly consisted of 9,10-epoxy ω-hydroxy and 9,10,ω-trihydroxy C18 alkanoic acids. The main aliphatic cutan monomers were alkanoic acids, primary alkanols, ω-hydroxy alkanoic acids and alkane-α,ω-diols ranging predominantly from C28 to C34 and maximizing at C32. Minor contributions of benzene-1,3,5-triol and derivatives suggested that these aromatic moieties form the polymeric core of cutan, to which the aliphatic moieties are linked via ester and possibly ether bonds. Conclusions High similarity of aliphatic moieties in the cutan and the cuticular wax component indicated a common biosynthetic origin. In order to exclude species-specific peculiarities of A. americana and to place our results in a broader context, cuticular waxes, cutin and cutan of Clivia miniata , Ficus elastica and Prunus laurocerasus leaves were also investigated. A detailed comparison showed compositional and structural differences, indicated that cutan was only found in leaves of perennial evergreen A. americana and C. miniata , and made clear that the phenomenon of cutan is possibly less present in plant species than suggested in the literature. [ABSTRACT FROM AUTHOR]

Wheat and flax crops are widely available and responsible for tons of straw generation every year around the world. These straws are by-products with excellent characteristics to industrial sectors, such as chemical, paper, textile, and biofuel. On the other hand, difficulties on storage and transportation of the straw, together with complex industrial processes and low profits from the final products, make the straw commerce unattractive. One possibility to increase the market value of straw is the recovery of vegetal waxes from them, which could be done without significant alterations on their structure, allowing them to be used further. Vegetal waxes have been intensively studied during the last few years by researchers in food, pharmaceutical, and chemical areas due to its organogelation capacity. The aim of this work is to present a comprehensive review starting from wheat and flax straw structure, passing through wax extraction, and finishing at the possible applications of wax compounds. Attention will be drawn to greener processing alternatives and technologies to selectively separate epicuticular waxes. [ABSTRACT FROM AUTHOR]

BACKGROUND Cherry fruit cracking is a costly problem for cherry growers. The effect of repeated sprayings (gibberellic acid – GA3; abscisic acid – ABA; salicylic acid – SA; glycine betaine – GB, and Ascophyllum nodosum – AN) combined with CaCl2, on 'Sweetheart' cherry fruit‐cracking characteristics was investigated. Cracking was quantified in terms of cracking incidence, crack morphology, confocal scanning laser microscopy, cuticular wax content, cell‐wall modification, and cuticular wax gene expression. RESULTS: All spray treatments reduced cracking compared with an untreated control (H2O), with fewer cheek cracks. The least cracking incidence was observed for ABA + CaCl2‐ and GB + CaCl2‐treated fruits, indicating an added benefit compared to spraying with CaCl2 alone. In addition, GB + CaCl2‐treated fruits showed higher fruit diameter. ABA + CaCl2 and GB + CaCl2 sprays showed higher wax content and higher cuticle and epidermal thickness compared with the control, including increased expression of wax synthase (ABA + CaCl2) and expansin 1 (GB + CaCl2). CONCLUSION: In general, factors that improve the cuticle thickness appear to be important at the fruit‐coloring stage. At the fruit‐ripening stage, larger cell sizes of the epidermis, hypodermis, and parenchyma cells lower cracking incidence, indicating the importance of flexibility and elasticity of the epidermis. © 2020 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Six perennial herbs (Plantago asiatica, Polygonum viviparum, Anaphalis lactea, Kobresia humilis, Leontopodium nanum and Potentilla chinensis) widely distributed in alpine meadows were reciprocally transplanted at two sites in eastern edge of Qinghai-Tibetan Plateau, Hongyuan (3434 m, 2.97 °C, 911 mm) and Qilian (3701 m, 2.52 °C, 472 mm), aiming to evaluate the responses of alpine plants to changing environments. When plants were transplanted from Hongyuan to Qilian, most plant species showed a decrease of total wax coverage in first year and reverse trend was observed for some plant species in second year. However, when plants were transplanted from Qilian to Hongyuan, the response of total wax coverage differed greatly between plant species. When compared with those in first year, plasticity index of average chain length of alkane decreased whereas carbon preference index of alkane increased at both Hongyuan and Qilian in second year. The total wax coverage differed between local and transplanted plants, suggesting both environmental and genetic factors controlled the wax depositions. Structural equation modeling indicated that co-variations existed between leaf cuticular waxes and leaf functional traits. These results suggest that alpine herbs adjust both wax depositions and chain length distributions to adapt to changing environment, showing climate adaptations.

Abstract Background While most water loss from leaf surfaces occurs via stomata, part of this loss also occurs through the leaf cuticle, even when the stomata are fully closed. This component, termed residual transpiration, dominates during the night and also becomes critical under stress conditions such as drought or salinity. Reducing residual transpiration might therefore be a potentially useful mechanism for improving plant performance when water availability is reduced (e.g. under saline or drought stress conditions). One way of reducing residual transpiration may be via increased accumulation of waxes on the surface of leaf. Residual transpiration and wax constituents may vary with leaf age and position as well as between genotypes. This study used barley genotypes contrasting in salinity stress tolerance to evaluate the contribution of residual transpiration to the overall salt tolerance, and also investigated what role cuticular waxes play in this process. Leaves of three different positions (old, intermediate and young) were used. Results Our results show that residual transpiration was higher in old leaves than the young flag leaves, correlated negatively with the osmolality, and was positively associated with the osmotic and leaf water potentials. Salt tolerant varieties transpired more water than the sensitive variety under normal growth conditions. Cuticular waxes on barley leaves were dominated by primary alcohols (84.7–86.9%) and also included aldehydes (8.90–10.1%), n-alkanes (1.31–1.77%), benzoate esters (0.44–0.52%), phytol related compounds (0.22–0.53%), fatty acid methyl esters (0.14–0.33%), β-diketones (0.07–0.23%) and alkylresorcinols (1.65–3.58%). A significant negative correlation was found between residual transpiration and total wax content, and residual transpiration correlated significantly with the amount of primary alcohols. Conclusions Both leaf osmolality and the amount of total cuticular wax are involved in controlling cuticular water loss from barley leaves under well irrigated conditions. A significant and negative relationship between the amount of primary alcohols and a residual transpiration implies that some cuticular wax constituents act as a water barrier on plant leaf surface and thus contribute to salinity stress tolerance. It is suggested that residual transpiration could be a fundamental mechanism by which plants optimize water use efficiency under stress conditions.

Cuticular waxes, long-chain hydrocarbon compounds, form the outermost layer of plant surfaces in most terrestrial plants. The presence of cuticular waxes protects plants from water loss and other environmental stresses. Cloning and characterization of genes involved in the regulation, biosynthesis, and extracellular transport of cuticular waxes onto the surface of epidermal cells have revealed the molecular basis of cuticular wax accumulation. However, intracellular trafficking of synthesized waxes to the plasma membrane for cellular secretion is poorly understood. Here, we characterized a maize glossy (gl6) mutant that exhibited decreased epicuticular wax load, increased cuticle permeability, and reduced seedling drought tolerance relative to wild-type. We combined an RNA-sequencing-based mapping approach (BSR-Seq) and chromosome walking to identify the gl6 candidate gene, which was confirmed via the analysis of multiple independent mutant alleles. The gl6 gene represents a novel maize glossy gene containing a conserved, but uncharacterized, DUF538 domain. This study suggests that the GL6 protein may be involved in the intracellular trafficking of cuticular waxes, opening the door to elucidating the poorly understood process by which cuticular wax is transported from its site of biosynthesis to the plasma membrane. [ABSTRACT FROM AUTHOR]

Waxes have an important protective role in the postharvest storage potential of fruit. However, there are few studies in which the waxes of fruit of kumquat cultivars have been analysed. In this work, the fruit of a kumquat mutant with a relatively thin skin ('Huapi' kumquat, HPK) was found to present lower fruit weight loss and decay during cold storage than the fruit of the control ('Rongan' kumquat, RAK) did. Scanning electron microscopy observations revealed the epicuticular wax crystals on the HPK fruit were more abundant and larger than those on the RAK fruit at harvest. Gas chromatography-mass spectrometry analysis showed the amounts of total wax, triterpenoids and most of the aliphatic wax fractions on the HPK fruit were much higher than those on the RAK fruit throughout the cold storage process. Correlation analysis showed that differences in amounts of fatty acids and aldehydes might be key factors responsible for the differences in the postharvest traits between the two kumquat cultivars. Additionally, the real-time quantitative RT-PCR analysis results indicated that 12 genes involved in wax synthesis, transport and transcription were considered to play important roles in regulating the wax formation of kumquat fruit during cold storage. • HPK presented lower fruit weight loss and decay than RAK during cold storage. • There were differences in wax morphology and content between HPK and RAK fruit. • Fatty acids involved in fruit decay in kumquat during cold storage. • Aldehydes involved in fruit weight loss and decay in kumquat during cold storage. • LACS1 , ACC1 , KCS6 and CER3 genes might related to the difference of storage ability. [ABSTRACT FROM AUTHOR]

The link between the MtTdp1α (tyrosyl-DNA phosphodiesterase) gene, involved in the repair of DNA topoisomerase I mediated DNA damage, and the plant defense response has been investigated in MtTdp1α-depleted Medicago truncatula transgenic lines obtained by intron-spliced hairpin RNA approach, compared to the control line (CTRL, empty vector). Reduction of cuticle permeability highlighted by chlorophyll efflux assays positively correlated with the level of MtTdp1α gene silencing. The increased cuticle thickness was confirmed by transmission electron microscopy, which revealed an apparent expansion of the epicuticular waxes deposited on the outer surface. RNA-Seq analysis, carried out in the MtTdp1α-depleted plants, revealed the different expression of resistance (R) genes, PAMP (pathogen-associated-molecular pattern) triggered immunity (PTI) genes and transcription factors (TFs) involved in the regulation of the plant defense response.

Austropuccinia psidii, the causal agent of myrtle rust, is a biotrophic pathogen whose growth and development depends on the host tissues. The uredospores of A. psidii infect Eucalyptus by engaging in close contact with the host surface and interacting with the leaf cuticle that provides important chemical and physical signals to trigger the infection process. In this study, the cuticular waxes of Eucalyptus spp. were analyzed to determine their composition or structure and correlation with susceptibility/resistance to A. psidii. Twenty-one Eucalyptus spp. in the field were classified as resistant or susceptible. The resistance/susceptibility level of six Eucalyptus spp. were validated in controlled conditions using qPCR, revealing that the pathogen can germinate on the eucalyptus surface of some species without multiplying in the host. CG-TOF-MS analysis detected 26 compounds in the Eucalyptus spp. cuticle and led to the discovery of the role of hexadecanoic acid in the susceptibility of Eucalyptus grandis and Eucalyptus phaeotricha to A. psidii. We characterized the epicuticular wax morphology of the six previously selected Eucalyptus spp. using scanning electron microscopy and observed different behavior in A. psidii germination during host infection. It was found a correlation of epicuticular morphology on the resistance to A. psidii. However, in this study, we provide the first report of considerable interspecific variation in Eucalyptus spp. on the susceptibility to A. psidii and its correlation with cuticular waxes chemical compounds that seem to play a synergistic role as a preformed defense mechanism.

The cuticle is a hydrophobic structure that seals plant aerial surfaces from the surrounding environment. To better understand how cuticular wax composition changes over development, we conducted an untargeted screen of leaf surface lipids from black cottonwood ( Populus trichocarpa ). We observed major shifts to the lipid profile across development, from a phenolic and terpene-dominated profile in young leaves to an aliphatic wax-dominated profile in mature leaves. Contrary to the general pattern, levels of aliphatic cis -9-alkenes decreased in older leaves following their accumulation. A thorough examination revealed that the decrease in cis -9-alkenes was accompanied by a concomitant increase in aldehydes, one of them being the volatile compound nonanal. By applying exogenous alkenes to P. trichocarpa leaves, we show that unsaturated waxes in the cuticle undergo spontaneous oxidative cleavage to generate aldehydes and that this process occurs similarly in other alkene-accumulating systems such as balsam poplar ( Populus balsamifera ) leaves and corn ( Zea mays ) silk. Moreover, we show that the production of cuticular wax-derived compounds can be extended to other wax components. In bread wheat ( Triticum aestivum ), 9-hydroxy-14,16-hentriacontanedione likely decomposes to generate 2-heptadecanone and 7-octyloxepan-2-one (a caprolactone). These findings highlight an unusual route to the production of plant volatiles that are structurally encoded within cuticular wax precursors. These processes could play a role in modulating ecological interactions and open the possibility for engineering bioactive volatile compounds into plant waxes., Competing Interests: Competing interests statement:The authors declare no competing interest.

The efficacy of the cuticular transpiration barrier and its resistance to elevated temperatures are significantly higher in a typical water-saver than in a water-spender plant growing in hot desert. Water-saver and water-spender strategies are successful adaptations allowing plants to cope with the limitations of hot desert habitats. We investigated whether the efficacy of the cuticular transpiration barrier and its susceptibility to high temperatures are ecophysiological traits differentially developed in the water-spender Citrullus colocynthis and the water-saver Phoenix dactylifera. Minimum leaf conductance (g min) at 25 °C was six times lower in P. dactylifera (1.1×10–5 m s–1) than in C. colocynthis (6.9×10–5 m s–1). Additionally, g min in the range 25–50 °C did not change in P. dactylifera but increased by a factor of 3.2 in C. colocynthis. Arrhenius formalism applied to the C. colocynthis g min led to a biphasic graph with a steep increase at temperatures ≥35 °C, whereas for P. dactylifera the graph was linear over all temperatures. Leaf cuticular wax coverage amounted to 4.2±0.4 µg cm–2 for C. colocynthis and 29.4±4.2 µg cm–2 for P. dactylifera. In both species, waxes were mainly composed of very-long-chain aliphatics. Midpoints of the wax melting ranges of P. dactylifera and C. colocynthis were 80 °C and 73 °C, respectively. We conclude that in P. dactylifera a particular wax and cutin chemistry prevents the rise of g min at elevated temperatures. [ABSTRACT FROM AUTHOR]

Austropuccinia psidii , the causal agent of myrtle rust, is a biotrophic pathogen whose growth and development depends on the host tissues. The uredospores of A. psidii infect Eucalyptus by engaging in close contact with the host surface and interacting with the leaf cuticle that provides important chemical and physical signals to trigger the infection process. In this study, the cuticular waxes of Eucalyptus spp. were analyzed to determine their composition or structure and correlation with susceptibility/resistance to A. psidii. Twenty-one Eucalyptus spp. in the field were classified as resistant or susceptible. The resistance/susceptibility level of six Eucalyptus spp. were validated in controlled conditions using qPCR, revealing that the pathogen can germinate on the eucalyptus surface of some species without multiplying in the host. CG-TOF-MS analysis detected 26 compounds in the Eucalyptus spp. cuticle and led to the discovery of the role of hexadecanoic acid in the susceptibility of Eucalyptus grandis and Eucalyptus phaeotricha to A. psidii. We characterized the epicuticular wax morphology of the six previously selected Eucalyptus spp. using scanning electron microscopy and observed different behavior in A. psidii germination during host infection. It was found a correlation of epicuticular morphology on the resistance to A. psidii. However, in this study, we provide the first report of considerable interspecific variation in Eucalyptus spp. on the susceptibility to A. psidii and its correlation with cuticular waxes chemical compounds that seem to play a synergistic role as a preformed defense mechanism. [ABSTRACT FROM AUTHOR]

Background and Aims The cuticular waxes sealing plant surfaces against excessive water loss are complex mixtures of very-long-chain aliphatics, with compositions that vary widely between plant species. To help fill the gap in our knowledge about waxes of non-flowering plant taxa, and thus about the cuticle of ancestral land plants, this study provides comprehensive analyses of waxes on temperate fern species from five different families. Methods The wax mixtures on fronds of Pteridium aquilinum, Cryptogramma crispa, Polypodium glycyrrhiza, Polystichum munitum and Gymnocarpium dryopteris were analysed using gas chromatography–mass spectrometry for identification, and gas chromatography–flame ionization detection for quantification. Key Results The wax mixtures from all five fern species contained large amounts of C36–C54 alkyl esters, with species-specific homologue distributions. They were accompanied by minor amounts of fatty acids, primary alcohols, aldehydes and/or alkanes, whose chain length profiles also varied widely between species. In the frond wax of G. dryopteris, C27–C33 secondary alcohols and C27–C35 ketones with functional groups exclusively on even-numbered carbons (C-10 to C-16) were identified; these are characteristic structures similar to secondary alcohols and ketones in moss, gymnosperm and basal angiosperm waxes. The ferns had total wax amounts varying from 3.9 μg cm–2 on P. glycyrrhiza to 16.9 μg cm–2 on G. dryopteris, thus spanning a range comparable with that on leaves of flowering plants. Conclusions The characteristic compound class compositions indicate that all five fern species contain the full complement of wax biosynthesis enzymes previously described for the angiosperm arabidopsis. Based on the isomer profiles, we predict that each fern species, in contrast to arabidopsis, has multiple ester synthase enzymes, each with unique substrate specificities. [ABSTRACT FROM AUTHOR]

Background and Aims Brassinosteroids (BRs) are plant hormones involved in many developmental processes as well as in plant–environment interactions. Their role was investigated in this study through the analysis of lilliputian1-1 (lil1-1), a dwarf mutant impaired in BR biosynthesis in maize (Zea mays). Methods We isolated lil1-1 through transposon tagging in maize. The action of lil1 was investigated through morphological and genetic analysis. Moreover, by comparing lil1-1 mutant and wild-type individuals grown under drought stress, the effect of BR reduction on the response to drought stress was examined. Key Results lil1-1 is a novel allele of the brassinosteroid-deficient dwarf1 (brd1) gene, encoding a brassinosteroid C-6 oxidase. We show in this study that lil1 is epistatic to nana plant1 (na1), a BR gene involved in earlier steps of the pathway. The lill-1 mutation causes alteration in the root gravitropic response, leaf epidermal cell density, epicuticular wax deposition and seedling adaptation to water scarcity conditions. Conclusions Lack of active BR molecules in maize causes a pleiotropic effect on plant development and improves seedling tolerance of drought. BR-deficient maize mutants can thus be instrumental in unravelling novel mechanisms on which plant adaptations to abiotic stress are based. [ABSTRACT FROM AUTHOR]

Objective – to determine the differences in the morphological parameters of stomata and the component composition of cuticular waxes of silver linden (Tilia tomentosa Moench) leaves under conditions of lighting and shading in the crown of a tree. Material and methods. The sun-adapted and shadeadapted fully developed leaves of silver linden were selected as the test objects in our study. The component composition of the cuticular waxes was investigated by gas chromatography method, and stomata size and quantity values were determined on the epidermal imprints of the abaxial side of leaves. Results. The adaptive changes in leaves of the alien invasive plant species silver linden were established under conditions of increasing light intensity and temperature and reducing air humidity. Conclusion. The leaf surface area, leaf weight per unit area, density of stomata, and the content of the long chain components of the cuticular waxes increase, the length and the width of stomata decrease under conditions of lighting.

Cuticular waxes are primarily composed of two classes of lipids: compounds derived from very-long-chain fatty acids and isoprenoids, particularly triterpenoids and steroids. Isoprenoids can occur in cuticular waxes in high amounts, dominating the mixture of aliphatic long-chain hydrocarbons, while in other plants they are found in trace concentrations. Triterpenoids occurring in fruit cuticular waxes are of interest due to their potential role in the protection against biotic stresses, including pathogen infections, and their impact on the mechanical toughness of the fruit surface, maintaining fruit integrity, and post-harvest quality. The aim of the present study was the determination of the changes in the triterpenoid profile of the fruit cuticular waxes of four plant species bearing edible berries: Vaccinium myrtillus, V. vitis-idaea, and Arbutus unedo of the Ericaceae and the edible honeysuckle Lonicera caerulea of the Caprifoliaceae. Triterpenoids were identified and quantified by GC-MS/FID (gas chromatography-mass spectrometry/flame ionization detection) at three different phenological stages: young berries, berries at the onset of ripening, and mature berries. During fruit development and maturation, the triterpenoid content in cuticular waxes displayed species-specific patterns of changes. The steroid content seemed to be directly correlated with the developmental stage, with a very typical point of transition between growth and ripening being observed in all the fruit analyzed in this study.