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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]

Climate change may alter soil microbial communities and soil organic matter (SOM) composition. Soil carbon (C) cycling takes place over multiple time scales; therefore, long-term studies are essential to better understand the factors influencing C storage and help predict responses to climate change. To investigate this further, soils that were heated by 5 °C above ambient soil temperatures for 18 years were collected from the Barre Woods Soil Warming Study at the Harvard Forest Long-term Ecological Research site. This site consists of large 30 × 30 m plots (control or heated) where entire root systems are exposed to sustained warming conditions. Measurements included soil C and nitrogen concentrations, microbial biomass, and SOM chemistry using gas chromatography–mass spectrometry and solid-state 13C nuclear magnetic resonance spectroscopy. These complementary techniques provide a holistic overview of all SOM components and a comprehensive understanding of SOM composition at the molecular-level. Our results showed that soil C concentrations were not significantly altered with warming; however, various molecular-level alterations to SOM chemistry were observed. We found evidence for both enhanced SOM decomposition and increased above-ground plant inputs with long-term warming. We also noted shifts in microbial community composition while microbial biomass remained largely unchanged. These findings suggest that prolonged warming induced increased availability of preferred substrates, leading to shifts in the microbial community and SOM biogeochemistry. The observed increase in gram-positive bacteria indicated changes in substrate availability as gram-positive bacteria are often associated with the decomposition of complex organic matter, while gram-negative bacteria preferentially break down simpler organic compounds altering SOM composition over time. Our results also highlight that additional plant inputs do not effectively offset chronic warming-induced SOM decomposition in temperate forests. [ABSTRACT FROM AUTHOR]

The valorisation of food by-products is an important step towards sustainability in food production. Tomatoes constitute one of the most processed crops in the world (160 million tonnes of tomatoes are processed every year), of which 4% is waste. This translates to 6.4 million tonnes of tomato skins and seeds. Currently, this waste is composted or is used in the production of low-value animal feed; higher value can be achieved if this waste stream is re-appropriated for more advanced purposes. Plant cuticle is a membrane structure found on leaves and fruit, including tomatoes, and is mainly composed of cutin. The main function of plant cuticle is to limit water loss from the internal tissue of the plant. Cutin, which can be recovered from the tomato skins by pH shift extraction, has hydrophobic (water repellent) properties and is therefore an ideal raw material for the development of a novel water-resistant coating. In this study, biomass-based bioplastics were developed. Unfortunately, although these bioplastics have good mechanical properties, their hydrophilic nature results in poor water barrier properties. To mitigate this, a very effective water-resistant coating was formulated using the cutin extracted from tomato peels. The water vapour permeability rates of the bioplastics improved by 74% and the percentage swelling of the bioplastic improved by 84% when treated with the cutin coating. With physicochemical properties that can compete with petroleum-based plastics, these bioplastics have the potential to address the growing market demand for sustainable alternatives for food packaging. Using ingredients generated from by-products of the food processing industries (circular economy), the development of these bioplastics also addresses the UN's Sustainable Development Goal (SDG) 12, Sustainable Consumption and Production (SCP). [ABSTRACT FROM AUTHOR]

The valorisation of food by-products is an important step towards sustainability in food production. Tomatoes constitute one of the most processed crops in the world (160 million tonnes of tomatoes are processed every year), of which 4% is waste. This translates to 6.4 million tonnes of tomato skins and seeds. Currently, this waste is composted or is used in the production of low-value animal feed; higher value can be achieved if this waste stream is re-appropriated for more advanced purposes. Plant cuticle is a membrane structure found on leaves and fruit, including tomatoes, and is mainly composed of cutin. The main function of plant cuticle is to limit water loss from the internal tissue of the plant. Cutin, which can be recovered from the tomato skins by pH shift extraction, has hydrophobic (water repellent) properties and is therefore an ideal raw material for the development of a novel water-resistant coating. In this study, biomass-based bioplastics were developed. Unfortunately, although these bioplastics have good mechanical properties, their hydrophilic nature results in poor water barrier properties. To mitigate this, a very effective water-resistant coating was formulated using the cutin extracted from tomato peels. The water vapour permeability rates of the bioplastics improved by 74% and the percentage swelling of the bioplastic improved by 84% when treated with the cutin coating. With physicochemical properties that can compete with petroleum-based plastics, these bioplastics have the potential to address the growing market demand for sustainable alternatives for food packaging. Using ingredients generated from by-products of the food processing industries (circular economy), the development of these bioplastics also addresses the UN’s Sustainable Development Goal (SDG) 12, Sustainable Consumption and Production (SCP).

Abstract Sweet corn rapidly loses its sweetness, along with other nutrients, once it is harvested, which reduces its economic value. The present study was conducted to elucidate the regulatory mechanisms by which a rapid cooling slurry ice (SI) treatment followed by storage at 0 ± 0.5°C extends the postharvest quality of sweet corn relative to slower direct cooling at 0 ± 0.5°C. In this study, we found that SI treatment can maintain the appearance quality of sweet corn and inhibit grain shrinkage, respiration, and ethylene release. SI treatment delayed the aging of corn by regulating the expression levels of genes and the levels of metabolites. SI treatment also maintains its carbohydrate content, which is mainly related to starch and sucrose metabolism, glycolysis, citric acid cycle, pentose phosphate pathway, and cutin biosynthesis. The results provide a possible target for improving the postharvest quality of sweet corn.

Abstract Background Ideally, the barrier properties of a fruit’s cuticle persist throughout its development. This presents a challenge for strawberry fruit, with their rapid development and thin cuticles. The objective was to establish the developmental time course of cuticle deposition in strawberry fruit. Results Fruit mass and surface area increase rapidly, with peak growth rate coinciding with the onset of ripening. On a whole-fruit basis, the masses of cutin and wax increase but on a unit surface-area basis, they decrease. The decrease is associated with marked increases in elastic strain. The expressions of cuticle-associated genes involved in transcriptional regulation (FaSHN1, FaSHN2, FaSHN3), synthesis of cutin (FaLACS2, FaGPAT3) and wax (FaCER1, FaKCS10, FaKCR1), and those involved in transport of cutin monomers and wax constituents (FaABCG11, FaABCG32) decreased until maturity. The only exceptions were FaLACS6 and FaGPAT6 that are presumably involved in cutin synthesis, and FaCER1 involved in wax synthesis. This result was consistent across five strawberry cultivars. Strawberry cutin consists mainly of C16 and C18 monomers, plus minor amounts of C19, C20, C22 and C24 monomers, ω-hydroxy acids, dihydroxy acids, epoxy acids, primary alcohols, carboxylic acids and dicarboxylic acids. The most abundant monomer is 10,16-dihydroxyhexadecanoic acid. Waxes comprise mainly long-chain fatty acids C29 to C46, with smaller amounts of C16 to C28. Wax constituents are carboxylic acids, primary alcohols, alkanes, aldehydes, sterols and esters. Conclusion The downregulation of cuticle deposition during development accounts for the marked cuticular strain, for the associated microcracking, and for their high susceptibility to the disorders of water soaking and cracking.

The present work describes the synthesis of poly(butylene succinate) (PBSu)-cutin copolymers by the two-stage melt polycondensation method, esterification and polycondensation. Cutin was added in four different concentrations, 2.5, 5, 10, and 20 wt%, in respect to succinic acid. The obtained copolymers were studied using a variety of techniques such as Fourier transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), polarized light microscopy (PLM), as well as diffuse reflectance spectroscopy (DRS). A series of results, in agreement between different techniques, revealed the formation of PBSu-cutin interactions, confirming indirectly the successful in situ synthetic route of copolymers. DSC and XRD combined with PLM results provided indications that the crystallization temperature increases with the addition of small amounts of cutin and gradually decreases with increasing concentration. The crystallization process was easier and faster at 2.5%, 5%, and 10% concentrations, whereas at 20%, it was comparable to neat PBSu. The presence of cutin, in general, leads to the facilitated crystallizability of PBSu (direct effect), whereas a moderate drop in the glass transition temperature is recorded, the latter being an indirect effect of cutin via crystallization. The thermal stability improved in the copolymers compared to neat PBSu. Water contact angle measurements confirmed that the addition of cutin decreased the hydrophilicity. The local and segmental relaxation mapping is demonstrated for PBSu/cutin here for the first time. Enzymatic hydrolysis and soil degradation tests showed that, overall, cutin accelerated the decomposition of the polymers. The copolymers may be proven useful in several applications. [ABSTRACT FROM AUTHOR]

The cuticle is a hydrophobic coating of most aerial plant surfaces crucial for limiting non-stomatal water loss. Plant cuticles consist of the lipid polyester cutin and associated waxes with compositions varying widely between plant species and organs. Here, we aimed to provide a comparative analysis of the dark-glossy adaxial and pale-glaucous abaxial sides of Drimys winteri (Winteraceae) leaves. Scanning electron microscopy showed nanotubular wax crystals lining the entire abaxial side of the leaf (including stomatal pores), while the adaxial side had patches of mixed platelet/tubule crystals and smooth areas between them. Consecutive treatments for wax removal and cutin depolymerization revealed that the waxes were deposited on a cutin network with micron-scale cavities across the entire abaxial surface including the stomata pores, and on a microscopically smooth cutin surface on the adaxial side of the leaf. Gas chromatography coupled to mass spectrometry and flame ionization detection showed that the wax mixtures on both sides of the leaf were complex mixtures of very-long-chain compounds dominated by the secondary alcohol nonacosan-10-ol and alkanediols with one hydroxyl on C-10. It is therefore very likely that the characteristic tubular wax crystals of both leaf sides are formed by these alcohols and diols. Further secondary alcohols and alkanediols, as well as ketols and alkanetriols with one functional group on C-10, were identified based on mass spectral fragmentation patterns. The similarities between all these mid-chain-functionalized compounds suggest that they are derived from nonacosan-10-ol via regio-specific hydroxylation reactions, likely catalyzed by three P450-dependent monooxygenases with different regio-specificities. [ABSTRACT FROM AUTHOR]

Background: Ideally, the barrier properties of a fruit's cuticle persist throughout its development. This presents a challenge for strawberry fruit, with their rapid development and thin cuticles. The objective was to establish the developmental time course of cuticle deposition in strawberry fruit. Results: Fruit mass and surface area increase rapidly, with peak growth rate coinciding with the onset of ripening. On a whole-fruit basis, the masses of cutin and wax increase but on a unit surface-area basis, they decrease. The decrease is associated with marked increases in elastic strain. The expressions of cuticle-associated genes involved in transcriptional regulation (FaSHN1, FaSHN2, FaSHN3), synthesis of cutin (FaLACS2, FaGPAT3) and wax (FaCER1, FaKCS10, FaKCR1), and those involved in transport of cutin monomers and wax constituents (FaABCG11, FaABCG32) decreased until maturity. The only exceptions were FaLACS6 and FaGPAT6 that are presumably involved in cutin synthesis, and FaCER1 involved in wax synthesis. This result was consistent across five strawberry cultivars. Strawberry cutin consists mainly of C16 and C18 monomers, plus minor amounts of C19, C20, C22 and C24 monomers, ω-hydroxy acids, dihydroxy acids, epoxy acids, primary alcohols, carboxylic acids and dicarboxylic acids. The most abundant monomer is 10,16-dihydroxyhexadecanoic acid. Waxes comprise mainly long-chain fatty acids C29 to C46, with smaller amounts of C16 to C28. Wax constituents are carboxylic acids, primary alcohols, alkanes, aldehydes, sterols and esters. Conclusion: The downregulation of cuticle deposition during development accounts for the marked cuticular strain, for the associated microcracking, and for their high susceptibility to the disorders of water soaking and cracking. [ABSTRACT FROM AUTHOR]

Global climate change has altered forest productivity across the globe. Although extreme drought and temperature have lowered forest productivity in some regions, increased productivity has been observed in many forests over the last few decades due to increases in atmospheric carbon dioxide, temperature, and nitrogen (N) deposition. These factors can lead to changes in both the quantity and quality of litterfall and root inputs to soil. Additionally, few studies have investigated multifactorial treatments (e.g., detrital changes and N deposition), on soil carbon (C) sequestration. To investigate the long-term compositional changes to soil organic matter (SOM) in response to litter and N inputs, soil samples were collected from the University of Michigan Biological Station (UMBS) Detrital Input and Removal Treatment (DIRT) site after 15 years of experimental treatments. The samples were characterized using elemental analysis, targeted SOM compound analyses, nuclear magnetic resonance spectroscopy and microbial biomass and community composition measurements. The exclusions of C inputs (litter and/or roots) resulted in molecular-level biogeochemical changes, however, the soils at UMBS are seemingly more resistant to losses in soil C compared to other DIRT studies. Although the addition treatments (Double Litter, Double Wood, Double Litter + N, and N) led to increased soil C concentrations at UMBS, we found evidence for enhanced SOM decomposition occurring with litter additions. Notably, the observations made from the concurrent treatment of Double Litter + N demonstrated that the responses of individual treatments are not representative of simultaneous applications. Collectively, these results demonstrate that soil C biogeochemistry is sensitive to fluctuations in C and N deposition and overall, these processes are dictated by site-specific ecological properties. [ABSTRACT FROM AUTHOR]

This study primarily aimed to enhance the extraction of cutin from industrial tomato peel residues. Initially, the conventional extraction process was optimized using response surface methodology (RSM). Subsequently, high-pressure homogenization (HPH) was introduced to improve extraction efficiency and sustainability. The optimization process focused on determining the optimal conditions for conventional extraction via chemical hydrolysis, including temperature (100–130 °C), time (15–120 min), and NaOH concentration (1–3%). The optimized conditions, determined as 130 °C, 120 min, and 3% NaOH solution, yielded a maximum cutin extraction of 32.5%. Furthermore, the results indicated that applying HPH pre-treatment to tomato peels before alkaline hydrolysis significantly increased the cutin extraction yield, reaching 46.1%. This represents an approximately 42% increase compared to the conventional process. Importantly, HPH pre-treatment enabled cutin extraction under milder conditions using a 2% NaOH solution, reducing NaOH usage by 33%, while still achieving a substantial cutin yield of 45.6%. FT-IR analysis confirmed that cutin obtained via both conventional and HPH-assisted extraction exhibited similar chemical structures, indicating that the main chemical groups and structure of cutin remained unaltered by HPH treatment. Furthermore, cutin extracts from both conventional and HPH-assisted extraction demonstrated thermal stability up to approximately 200 °C, with less than 5% weight loss according to TGA analysis. These findings underscore the potential of HPH technology to significantly enhance cutin extraction yield from tomato peel residues while utilizing milder chemical hydrolysis conditions, thereby promoting a more sustainable and efficient cutin extraction process. [ABSTRACT FROM AUTHOR]

A recent study published in the Journal of Experimental Botany explores the role of NAC transcription factors in cuticle development in tomato fruit. The researchers found that the NAC TF SlNOR-like1 has a dual function in cuticle development, promoting cutin deposition and thickening the outer layer of the fruit, while also inhibiting wax accumulation. These findings provide new insights into the regulation of cuticle development in fruit and offer potential targets for improving fruit quality and storability through biotechnological approaches. The study highlights the importance of further characterizing the remaining "orphan" NAC TF family members to better understand their biological functions. [Extracted from the article]

The plant cuticle is an important protective barrier on the plant surface, constructed mainly by polymerized cutin matrix and a complex wax mixture. Although the pathway of plant cuticle biosynthesis has been clarified, knowledge of the transcriptional regulation network underlying fruit cuticle formation remains limited. In the present work, we discovered that tomato fruits of the NAC transcription factor SlNOR-like1 knockout mutants (nor-like1) produced by CRISPR/Cas9 [clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9] displayed reduced cutin deposition and cuticle thickness, with a microcracking phenotype, while wax accumulation was promoted. Further research revealed that SlNOR-like1 promotes cutin deposition by binding to the promoters of glycerol-3-phosphate acyltransferase6 (SlGPAT6 ; a key gene for cutin monomer formation) and CUTIN DEFICIENT2 (SlCD2 ; a positive regulator of cutin production) to activate their expression. Meanwhile, SlNOR-like1 inhibits wax accumulation, acting as a transcriptional repressor by targeting wax biosynthesis, and transport-related genes 3-ketoacyl-CoA synthase1 (SlKCS1), ECERIFERUM 1-2 (SlCER1-2), SlWAX2 , and glycosylphosphatidylinositol-anchored lipid transfer protein 1-like (SlLTPG1-like). In conclusion, SlNOR-like1 executes a dual regulatory effect on tomato fruit cuticle development. Our results provide a new model for the transcriptional regulation of fruit cuticle formation. [ABSTRACT FROM AUTHOR]

Cutin, one of the main structural components of tomato peels, is a waxy biopolymer rich in hydroxylated fatty acids. In this study, 10,16-dihydroxyhexadecanoic acid (10,16-diHHDA) was extracted and isolated from tomato peels and exploited to develop fully crosslinked polyesters as potential candidates for replacing fossil-based metal protective coatings. A preliminary screening was conducted to select the base formulation, and then a design of experiments (DoE) was used as a methodology to identify the optimal composition to develop a suitable coating material. Different formulations containing 10,16-diHHDA and other biorefinery monomers, including 2,5-furandicarboxylic acid, were considered. To this end, all polyesters were characterized through differential scanning calorimetry (DSC) and gel content measurements to determine their Tg value and crosslinking efficiency. Compositions exhibiting the best trade-off between Tg value, chemical resistance, and sufficiently high 10,16-diHHDA content between 39 and 48 wt.% were used to prepare model coatings that were characterized for assessing their wettability, scratch hardness, chemical resistance, and adhesion to metal substrates. These polyester coatings showed a Tg in the range of 45–55 °C, a hydrophobic behavior with a water contact angle of around 100°, a good solvent resistance (>100 MEK double rubs), and an adhesion strength to steel higher than 2 MPa. The results obtained confirmed the potential of cutin-based resins as coatings for metal protection, meeting the requirements for ensuring physicochemical properties of the final product, as well as for optimizing the valorization of such an abundant agri-food waste as tomato peels. [ABSTRACT FROM AUTHOR]

Pine nut skin (Pinus pinea L.) is a poorly explored industrial byproduct with potential to be utilized as a food ingredient. Subcritical water extraction (SWE), an eco-friendly extraction technique with higher efficiency than hot-water extraction (HWE), was studied to evaluate its suitability in producing extracts rich in soluble fiber and phenolic compounds. For this, a factorial design was developed considering temperature (120–180 °C), time (2–10 min), and mass/volume ratio (1–3 g/60 mL) under microwave irradiation. This design aimed to maximize the extraction of carbohydrates, while achieving the highest content of phenolic compounds and antioxidant activity. SWE produced higher yields (2.6-fold in relation to HWE) of extracts rich in polysaccharides, determined by methylation analysis, oligosaccharides, determined by GC-qMS as alditol acetates, and phenolic compounds, determined by HPLC-ESI-MS. SWE increased the recovery of pectic oligosaccharides (10-fold) and xyloglucans (2-fold), and allowed to recover pectic polysaccharides, type II arabinogalactans and insoluble-bound phenolic compounds. Mono-, oligo- and polysaccharides were not hydrolyzed during in vitro gastrointestinal digestion, showing potential prebiotic functionality. Although phenolic compounds suffered a 23 % (gallic acid equivalents) decrease, phenolic acids and aldehydes were released or conserved upon intestinal phase. These results highlight the potential of PNS valorization as functional food ingredients through the subcritical water solubilization of polysaccharides and phenolic compounds.

Cellular damage infiicted by wounding, pathogen infection, and herbivory releases a variety of host-derived metabolites, degraded structural components, and peptides into the extracellular space that act as alarm signals when perceived by adjacent cells. These so-called damage-associated molecular patterns (DAMPs) function through plasma membrane localized pattern recognition receptors to regulate wound and immune responses. In plants, DAMPs act as elicitors themselves, often inducing immune outputs such as calcium influx, reactive oxygen species generation, defense gene expression, and phytohormone signaling. Consequently, DAMP perception results in a priming effect that enhances resistance against subsequent pathogen in fections. Alongside their established function in local tissues, recent evidence supports a critical role of DAMP signaling in generation and/or amplification of mobile signals that induce systemic immune priming. Here, we summarize the identity, signaling, and synergy of proposed and established plant DAMPs. with a focus on those with published roles in systemic signaling. [ABSTRACT FROM AUTHOR]

The outermost hydrophobic layer of plants, i.e. the cuticle, is mainly composed of cutin, a polyester of hydroxy fatty acids with reported eliciting and/or antimicrobial activities for some of them. By-products of the fruit processing industry (fruit pomaces), often strongly enriched in cuticular material, are therefore a potential source of bioactive compounds for crop protection against pathogen attack. We investigated the utilization of tomato and apple pomaces in the development of a cutin-based biocontrol solution against apple scab, a major apple disease caused by Venturia inaequalis. Several cutin monomer extracts obtained through different strategies of depolymerization and purification were first compared for their ability to induce a targeted set of defense genes in apple seedlings after foliar application. After a step of formulation, some extracts were chosen for further investigation in planta and in vitro. Our results show that formulated cutin monomers could trigger a significant transcriptome reprogramming in apple plants and exhibit an antifungal effect on V. inaequalis. Cutin monomers-treated apple seedlings were significantly protected against infection by the apple scab agent. Altogether, our findings suggest that water-dispersed cutin monomers extracted from pomaces are potential new bio-based solutions for the control of apple scab. [ABSTRACT FROM AUTHOR]

This study focuses on the role of the qualitative leaf wax composition in modulating the cuticular water loss using a Populus × canescens cer6 mutant line, which accumulates C34–C46 wax ester dimers and is reduced in wax monomers >C24. The two literature‐based hypotheses to be tested were the importance of the amount of wax esters and the weighted mean carbon chain length in restricting cuticular water loss. The main results were acquired by chemical analysis of cuticular wax and gravimetric cuticular transpiration measurements. Besides additional physiological measurements, the leaf surface properties were characterised by scanning electron microscopy and spectrophotometric light reflectance quantification. Mutation of the CER6 gene resulted in striking changes in qualitative wax composition but not quantitative wax amount. Based on the strong accumulation of dimeric wax esters, the relative proportion of esters increased to >90%, and the weighted mean carbon chain length increased by >6 carbon atoms. These qualitative alterations were found to increase the cuticular transpiration of leaves by twofold. Our results do not support the hypotheses that enhanced amounts of wax esters or increased weighted mean carbon chain lengths of waxes lead to reduced cuticular transpiration. Summary statement: Mutation of the CER6 gene in Populus × canescens significantly increased the proportion of wax esters and the weighted mean carbon chain length of the cuticular wax. However, in contrast to literature‐based expectations, these alterations increased the cuticular transpiration by twofold. [ABSTRACT FROM AUTHOR]

Green tomato (Physalis ixocarpa) is a specie native to Mexico, and it is known as "tomatillo" or "husk tomato". The fruit contains vitamins, minerals, phenolic compounds, and steroidal lactones, presenting antimicrobial activity and antinarcotic effects. Therefore, it is not only used in traditional Mexican cuisine, but also in traditional medicine to relieve some discomforts such as fever, cough, and amygdalitis. However, it is a perishable fruit whose shelf life is very short. As a part of the peel, cuticle, and epicuticular waxes represent the most important part in plant protection, and the specific composition and structural characterization are significant to know how this protective biopolymer keeps quality characteristics in fresh fruits. P. ixocarpa cutin was obtained by enzymatic treatments (cellulase, hemicellulose, and pectinase) and different concentrations of TFA, and studied through Cross Polarization Magic Angle Spinning Nuclear Magnetic Resonance (CPMAS 13C NMR), Ultra-High Performance Liquid Chromatography coupled to Mass Spectrometry (UHPLC-MS), and was morphologically characterized by Confocal Laser Scanning Microscopy (CLSM) and Scanning Electron Microscopy (SEM). The main constituents identified under the basis of UHPLC-MS analysis were 9,10,18-trihydroxy-octadecanoic acid and 9,10-epoxy-18-hydroxy-octadecanoic acid with 44.7 and 37.5%, respectively. The C16 absence and low occurrence of phenolic compounds, besides the presence of glandular trichomes, which do not allow a continuous layer on the surface of the fruit, could be related to a lower shelf life compared with other common fruits such as tomato (Solanum lycopersicum). [ABSTRACT FROM AUTHOR]

The present work describes the synthesis of poly(butylene succinate) (PBSu)-cutin copolymers by the two-stage melt polycondensation method, esterification and polycondensation. Cutin was added in four different concentrations, 2.5, 5, 10, and 20 wt%, in respect to succinic acid. The obtained copolymers were studied using a variety of techniques such as Fourier transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), polarized light microscopy (PLM), as well as diffuse reflectance spectroscopy (DRS). A series of results, in agreement between different techniques, revealed the formation of PBSu-cutin interactions, confirming indirectly the successful in situ synthetic route of copolymers. DSC and XRD combined with PLM results provided indications that the crystallization temperature increases with the addition of small amounts of cutin and gradually decreases with increasing concentration. The crystallization process was easier and faster at 2.5%, 5%, and 10% concentrations, whereas at 20%, it was comparable to neat PBSu. The presence of cutin, in general, leads to the facilitated crystallizability of PBSu (direct effect), whereas a moderate drop in the glass transition temperature is recorded, the latter being an indirect effect of cutin via crystallization. The thermal stability improved in the copolymers compared to neat PBSu. Water contact angle measurements confirmed that the addition of cutin decreased the hydrophilicity. The local and segmental relaxation mapping is demonstrated for PBSu/cutin here for the first time. Enzymatic hydrolysis and soil degradation tests showed that, overall, cutin accelerated the decomposition of the polymers. The copolymers may be proven useful in several applications.

The outermost hydrophobic layer of plants, i.e. the cuticle, is mainly composed of cutin, a polyester of hydroxy fatty acids with reported eliciting and/or antimicrobial activities for some of them. By-products of the fruit processing industry (fruit pomaces), often strongly enriched in cuticular material, are therefore a potential source of bioactive compounds for crop protection against pathogen attack. We investigated the utilization of tomato and apple pomaces in the development of a cutin-based biocontrol solution against apple scab, a major apple disease caused by Venturia inaequalis. Several cutin monomer extracts obtained through different strategies of depolymerization and purification were first compared for their ability to induce a targeted set of defense genes in apple seedlings after foliar application. After a step of formulation, some extracts were chosen for further investigation in planta and in vitro. Our results show that formulated cutin monomers could trigger a significant transcriptome reprogramming in apple plants and exhibit an antifungal effect on V. inaequalis. Cutin monomers-treated apple seedlings were significantly protected against infection by the apple scab agent. Altogether, our findings suggest that water-dispersed cutin monomers extracted from pomaces are potential new bio-based solutions for the control of apple scab.

The plant cuticle, consisting of wax and cutin, is involved in adaptations to various environments. β-Ketoacyl-CoA synthases (KCSs) usually serve as a component of the fatty acid elongation complex that participates in the production of very long-chain fatty acids and provides precursors for the synthesis of various lipids, including wax; however, we recently reported that KCS3 and KCS12 negatively regulate wax biosynthesis. In this current study, we observed that unlike KCS3 -overexpressing (OE) lines, KCS12 -OE lines had fused floral organs because of abnormal cuticle biosynthesis. This prompted us to compare the functions of KCS3 and KCS12 during cuticle formation. Mutation of KCS3 caused greater effects on wax production, whereas mutation of KCS12 exerted more severe effects on cutin synthesis. The double-mutant kcs3 kcs12 had significantly increased wax and cutin contents compared to either single-mutant, suggesting that KCS12 and KCS3 have additive effects on cuticle biosynthesis. Cuticle permeability was greater for the double-mutant than for the single mutants, which ultimately led to increased susceptibility to drought stress and floral-organ fusion. Taken together, our results demonstrate the regulatory roles of KCS3 and KCS12 during cuticle biosynthesis, and show that maintaining KCS3 and KCS12 expression at certain levels is essential for the formation of a functional cuticle layer. [ABSTRACT FROM AUTHOR]

Summary: The cuticle is a protective layer covering aerial plant organs. We studied the function of waxes for the establishment of the cuticular barrier in barley (Hordeum vulgare). The barley eceriferum mutants cer‐za.227 and cer‐ye.267 display reduced wax loads, but the genes affected, and the consequences of the wax changes for the barrier function remained unknown.Cuticular waxes and permeabilities were measured in cer‐za.227 and cer‐ye.267. The mutant loci were isolated by bulked segregant RNA sequencing. New cer‐za alleles were generated by genome editing. The CER‐ZA protein was characterized after expression in yeast and Arabidopsis cer4‐3.Cer‐za.227 carries a mutation in HORVU5Hr1G089230 encoding acyl‐CoA reductase (FAR1). The cer‐ye.267 mutation is located to HORVU4Hr1G063420 encoding β‐ketoacyl‐CoA synthase (KAS1) and is allelic to cer‐zh.54. The amounts of intracuticular waxes were strongly decreased in cer‐ye.267. The cuticular water loss and permeability of cer‐za.227 were similar to wild‐type (WT), but were increased in cer‐ye.267. Removal of epicuticular waxes revealed that intracuticular, but not epicuticular waxes are required to regulate cuticular transpiration.The differential decrease in intracuticular waxes between cer‐za.227 and cer‐ye.267, and the removal of epicuticular waxes indicate that the cuticular barrier function mostly depends on the presence of intracuticular waxes. [ABSTRACT FROM AUTHOR]

This study primarily aimed to enhance the extraction of cutin from industrial tomato peel residues. Initially, the conventional extraction process was optimized using response surface methodology (RSM). Subsequently, high-pressure homogenization (HPH) was introduced to improve extraction efficiency and sustainability. The optimization process focused on determining the optimal conditions for conventional extraction via chemical hydrolysis, including temperature (100–130 °C), time (15–120 min), and NaOH concentration (1–3%). The optimized conditions, determined as 130 °C, 120 min, and 3% NaOH solution, yielded a maximum cutin extraction of 32.5%. Furthermore, the results indicated that applying HPH pre-treatment to tomato peels before alkaline hydrolysis significantly increased the cutin extraction yield, reaching 46.1%. This represents an approximately 42% increase compared to the conventional process. Importantly, HPH pre-treatment enabled cutin extraction under milder conditions using a 2% NaOH solution, reducing NaOH usage by 33%, while still achieving a substantial cutin yield of 45.6%. FT-IR analysis confirmed that cutin obtained via both conventional and HPH-assisted extraction exhibited similar chemical structures, indicating that the main chemical groups and structure of cutin remained unaltered by HPH treatment. Furthermore, cutin extracts from both conventional and HPH-assisted extraction demonstrated thermal stability up to approximately 200 °C, with less than 5% weight loss according to TGA analysis. These findings underscore the potential of HPH technology to significantly enhance cutin extraction yield from tomato peel residues while utilizing milder chemical hydrolysis conditions, thereby promoting a more sustainable and efficient cutin extraction process.

Cutin, one of the main structural components of tomato peels, is a waxy biopolymer rich in hydroxylated fatty acids. In this study, 10,16-dihydroxyhexadecanoic acid (10,16-diHHDA) was extracted and isolated from tomato peels and exploited to develop fully crosslinked polyesters as potential candidates for replacing fossil-based metal protective coatings. A preliminary screening was conducted to select the base formulation, and then a design of experiments (DoE) was used as a methodology to identify the optimal composition to develop a suitable coating material. Different formulations containing 10,16-diHHDA and other biorefinery monomers, including 2,5-furandicarboxylic acid, were considered. To this end, all polyesters were characterized through differential scanning calorimetry (DSC) and gel content measurements to determine their Tg value and crosslinking efficiency. Compositions exhibiting the best trade-off between Tg value, chemical resistance, and sufficiently high 10,16-diHHDA content between 39 and 48 wt.% were used to prepare model coatings that were characterized for assessing their wettability, scratch hardness, chemical resistance, and adhesion to metal substrates. These polyester coatings showed a Tg in the range of 45–55 °C, a hydrophobic behavior with a water contact angle of around 100°, a good solvent resistance (>100 MEK double rubs), and an adhesion strength to steel higher than 2 MPa. The results obtained confirmed the potential of cutin-based resins as coatings for metal protection, meeting the requirements for ensuring physicochemical properties of the final product, as well as for optimizing the valorization of such an abundant agri-food waste as tomato peels.

Polylactic acid (PLA) is one of the most important biopolymers employed on the market due to its good mechanical strength and barrier properties. On the other hand, this material presents a rather low flexibility, limiting its employment. The valorization of bio-based agro-food waste for the modification of bioplastics is a highly appealing approach for the replacement of petrol-based materials. The aim of this work is to employ cutin fatty acids derived from a biopolymer (i.e., cutin), present in waste tomato peels and its bio-based derivatives as new plasticizers to enhance PLA flexibility. In particular, pure 10,16-dihydroxy hexadecanoic acid was extracted and isolated from tomato peels and then functionalized to give the desired compounds. All the molecules developed in this study were characterized by NMR and ESI-MS. Blends at different concentrations (10, 20, 30, and 40% w/w) the flexibility (Tg measurements with differential scanning calorimetry—DSC) of the final material. Furthermore, the physical behavior of two blends obtained by mechanical mixing of PLA and 16-methoxy,16-oxohexadecane-1,7-diyl diacetate was investigated through thermal and tensile tests. The data collected by DSC show a lowering in the Tg of all the blends of PLA with functionalized fatty acids, in comparison with pure PLA. Lastly, the tensile tests highlighted how PLA blended with 16-methoxy,16-oxohexadecane-1,7-diyl diacetate (20% w/w) can efficiently enhance its flexibility. [ABSTRACT FROM AUTHOR]

Summary: The plant cuticle is an important plant‐atmosphere boundary, the synthesis and maintenance of which represents a significant metabolic cost. Only limited information regarding cuticle dynamics is available.We determined the composition and dynamics of Clusia rosea cuticular waxes and matrix using 13CO2 labelling, compound‐specific and bulk isotope ratio mass spectrometry. Collodion was used for wax collection; gas exchange techniques to test for any collodion effects on living leaves.Cutin matrix (MX) area density did not vary between young and mature leaves and between leaf sides. Only young leaves incorporated new carbon into their MX. Collodion‐based sampling discriminated between epicuticular (EW) and intracuticular wax (IW) effectively. Epicuticular differed in composition from IW. The newly synthetised wax was deposited in IW first and later in EW. Both young and mature leaves synthetised IW and EW. The faster dynamics in young leaves were due to lower wax coverage, not a faster synthesis rate. Longer‐chain alkanes were deposited preferentially on the abaxial, stomatous leaf side, producing differences between leaf sides in wax composition.We introduce a new, sensitive isotope labelling method and demonstrate that cuticular wax is renewed during leaf ontogeny of C. rosea. We discuss the ecophysiological significance of the new insights. [ABSTRACT FROM AUTHOR]

Olive (Olea europaea L.) fruit and derived products play a pivotal role in the Mediterranean diet, to which they contribute their gastronomic value and their health-promoting properties. The fruit cuticle constitutes the interface between the plant and the surrounding environment, and it modulates relevant traits such as water loss, mechanical resistance, and susceptibility to pests and rots. Hence, a better knowledge of fruit cuticle properties and the impact thereupon of agronomic factors could help improving olive grove management. In this work, time–course changes in fruit cuticle yields and composition were assessed during the on-tree ripening of 'Arbequina' olives obtained from irrigated or rain-fed trees grown at a commercial grove located in El Soleràs (Catalonia, Spain), where low annual rainfall occur together with cold winters and hot dry summers. Significantly higher wax contents were observed for rain-fed than for irrigated fruits, both in relative (% over total cuticle) and in absolute terms (from 231 to 840 µg cm−2 and from 212 to 560 µg cm−2, respectively, contingent upon the maturity stage), in agreement with their proposed role as a barrier against water loss. Compositional differences in cuticular waxes and in cutin monomers were also detected between irrigated and rain-fed olives, with major changes involving significantly higher loads per surface area of triterpenoids and ω-hydroxy fatty acids in the latter. In contrast to the load and composition of cuticular wax, no apparent impact of irrigation was observed on either total cuticle yields or cuticle thickness. [ABSTRACT FROM AUTHOR]

Green tomato (Physalis ixocarpa) is a specie native to Mexico, and it is known as “tomatillo” or “husk tomato”. The fruit contains vitamins, minerals, phenolic compounds, and steroidal lactones, presenting antimicrobial activity and antinarcotic effects. Therefore, it is not only used in traditional Mexican cuisine, but also in traditional medicine to relieve some discomforts such as fever, cough, and amygdalitis. However, it is a perishable fruit whose shelf life is very short. As a part of the peel, cuticle, and epicuticular waxes represent the most important part in plant protection, and the specific composition and structural characterization are significant to know how this protective biopolymer keeps quality characteristics in fresh fruits. P. ixocarpa cutin was obtained by enzymatic treatments (cellulase, hemicellulose, and pectinase) and different concentrations of TFA, and studied through Cross Polarization Magic Angle Spinning Nuclear Magnetic Resonance (CPMAS 13C NMR), Ultra-High Performance Liquid Chromatography coupled to Mass Spectrometry (UHPLC-MS), and was morphologically characterized by Confocal Laser Scanning Microscopy (CLSM) and Scanning Electron Microscopy (SEM). The main constituents identified under the basis of UHPLC-MS analysis were 9,10,18-trihydroxy-octadecanoic acid and 9,10-epoxy-18-hydroxy-octadecanoic acid with 44.7 and 37.5%, respectively. The C16 absence and low occurrence of phenolic compounds, besides the presence of glandular trichomes, which do not allow a continuous layer on the surface of the fruit, could be related to a lower shelf life compared with other common fruits such as tomato (Solanum lycopersicum).

Forest ecosystems are important global soil carbon (C) reservoirs, but their capacity to sequester C is susceptible to climate change factors that alter the quantity and quality of C inputs. To better understand forest soil C responses to altered C inputs, we integrated three molecular composition published data sets of soil organic matter (SOM) and soil microbial communities for mineral soils after 20 years of detrital input and removal treatments in two deciduous forests: Bousson Forest (BF), Harvard Forest (HF), and a coniferous forest: H.J. Andrews Forest (HJA). Soil C turnover times were estimated from radiocarbon measurements and compared with the molecular‐level data (based on nuclear magnetic resonance and specific analysis of plant‐ and microbial‐derived compounds) to better understand how ecosystem properties control soil C biogeochemistry and dynamics. Doubled aboveground litter additions did not increase soil C for any of the forests studied likely due to long‐term soil priming. The degree of SOM decomposition was higher for bacteria‐dominated sites with higher nitrogen (N) availability while lower for the N‐poor coniferous forest. Litter exclusions significantly decreased soil C, increased SOM decomposition state, and led to the adaptation of the microbial communities to changes in available substrates. Finally, although aboveground litter determined soil C dynamics and its molecular composition in the coniferous forest (HJA), belowground litter appeared to be more influential in broadleaf deciduous forests (BH and HF). This synthesis demonstrates that inherent ecosystem properties regulate how soil C dynamics change with litter manipulations at the molecular‐level. Across the forests studied, 20 years of litter additions did not enhance soil C content, whereas litter reductions negatively impacted soil C concentrations. These results indicate that soil C biogeochemistry at these temperate forests is highly sensitive to changes in litter deposition, which are a product of environmental change drivers. [ABSTRACT FROM AUTHOR]

Wheat and barley are widely distributed cereal crops whose yields are adversely affected by environmental stresses such as drought, salinity, extreme temperatures, and attacks of pathogens and pests. As the interphase between aerial plant organs and their environments, hydrophobic cuticle largely consists of a cutin matrix impregnated and sealed with cuticular waxes. Increasing evidence supports that the cuticle plays a key role in plant adaptation to abiotic and biotic stresses, which could be harnessed for wheat and barley improvement. In this review, we highlighted recent advances in cuticle biosynthesis and its multifaceted roles in abiotic and biotic stress tolerance of wheat and barley. Current strategies, challenges, and future perspectives on manipulating cuticle biosynthesis for abiotic and biotic stress tolerance in wheat and barley are discussed. [ABSTRACT FROM AUTHOR]

Global environmental change is altering the quality and quantity of plant inputs into soil. However, it is unclear how these long-term changes may fundamentally shift the biogeochemistry of soil carbon in forests. To better understand how varied detrital inputs alter soil organic matter (OM) biogeochemistry and composition at the molecular-level, soil samples were collected from a 20 year detrital manipulation experiment in an old-growth coniferous rainforest in Western Oregon. The experiment includes ambient (control) plots and six treatments: Double Litter, Double Wood, No Roots, No Litter, No Inputs and OA-less (replacement of O and A horizons with B horizon). Total soil carbon and nitrogen, molecular-level OM composition using solid-state 13C nuclear magnetic resonance, and targeted compound extractions were measured. Although soil carbon did not increase with Double Litter and Double Wood, microbial biomass and the decomposition of specific forms of soil OM (i.e., cellulose) increased, likely due to sustained soil priming over 20 years. Mineral (0–10 cm) soil carbon was similar across litter exclusion treatments (No Litter, No Roots, No Inputs), however, soil OM decomposition increased relative to the control. Microbial-derived lipids increased under Double Litter but decreased when above-ground litter was excluded. Notably, needle-derived lipids decreased with above-ground litter exclusion and root-derived compounds did not change under below-ground root exclusion. These results suggested that above-ground litter alters soil carbon biogeochemistry in surface soils to a greater extent than below-ground inputs. This study also demonstrated that long-term soil carbon biogeochemical trajectories were mostly governed by litter quality, quality and microbial processing of above-ground inputs. [ABSTRACT FROM AUTHOR]

Fresh blueberries are prone to softening and dehydration during postharvest, which limits their competitiveness when reaching the final markets. Commercial cultivars 'Duke' and 'Brigitta' exhibit contrasting softening patterns. Although 'Duke' berries usually show higher firmness levels at harvest as compared to 'Brigitta', they display higher softening and weight loss rates after cold storage. The aim of this study was to evaluate the physicochemical changes and modifications in cuticle composition of 'Duke' and 'Brigitta' blueberries across five developmental stages: green (G), 25 and 50% pink (25P, 50P), and 75 and 100% blue (75B, 100B), to determine those characters with the most influence on their postharvest behavior. For each developmental stage, maturity parameters, respiration, and ethylene production rates were assessed, and cuticular wax and cutin were analyzed. Principal component analysis (PCA) revealed that 'Duke' berries were characterized by higher respiration and ethylene production rates, while 'Brigitta' showed higher contents of oleanolic acid and α-amyrin over total waxes. The results suggest that larger surface/volume ratios and higher amounts of ursolic acid and lupeol in ripe fruit may underlie higher weight and firmness loss rates of 'Duke' berries as compared to 'Brigitta'. [ABSTRACT FROM AUTHOR]

Polylactic acid (PLA) is one of the most important biopolymers employed on the market due to its good mechanical strength and barrier properties. On the other hand, this material presents a rather low flexibility, limiting its employment. The valorization of bio-based agro-food waste for the modification of bioplastics is a highly appealing approach for the replacement of petrol-based materials. The aim of this work is to employ cutin fatty acids derived from a biopolymer (i.e., cutin), present in waste tomato peels and its bio-based derivatives as new plasticizers to enhance PLA flexibility. In particular, pure 10,16-dihydroxy hexadecanoic acid was extracted and isolated from tomato peels and then functionalized to give the desired compounds. All the molecules developed in this study were characterized by NMR and ESI-MS. Blends at different concentrations (10, 20, 30, and 40% w/w) the flexibility (Tg measurements with differential scanning calorimetry—DSC) of the final material. Furthermore, the physical behavior of two blends obtained by mechanical mixing of PLA and 16-methoxy,16-oxohexadecane-1,7-diyl diacetate was investigated through thermal and tensile tests. The data collected by DSC show a lowering in the Tg of all the blends of PLA with functionalized fatty acids, in comparison with pure PLA. Lastly, the tensile tests highlighted how PLA blended with 16-methoxy,16-oxohexadecane-1,7-diyl diacetate (20% w/w) can efficiently enhance its flexibility.

Olive (Olea europaea L.) fruit and derived products play a pivotal role in the Mediterranean diet, to which they contribute their gastronomic value and their health-promoting properties. The fruit cuticle constitutes the interface between the plant and the surrounding environment, and it modulates relevant traits such as water loss, mechanical resistance, and susceptibility to pests and rots. Hence, a better knowledge of fruit cuticle properties and the impact thereupon of agronomic factors could help improving olive grove management. In this work, time–course changes in fruit cuticle yields and composition were assessed during the on-tree ripening of ‘Arbequina’ olives obtained from irrigated or rain-fed trees grown at a commercial grove located in El Soleràs (Catalonia, Spain), where low annual rainfall occur together with cold winters and hot dry summers. Significantly higher wax contents were observed for rain-fed than for irrigated fruits, both in relative (% over total cuticle) and in absolute terms (from 231 to 840 µg cm−2 and from 212 to 560 µg cm−2, respectively, contingent upon the maturity stage), in agreement with their proposed role as a barrier against water loss. Compositional differences in cuticular waxes and in cutin monomers were also detected between irrigated and rain-fed olives, with major changes involving significantly higher loads per surface area of triterpenoids and ω-hydroxy fatty acids in the latter. In contrast to the load and composition of cuticular wax, no apparent impact of irrigation was observed on either total cuticle yields or cuticle thickness.

Fleshy fruits represent a valuable resource of economic and nutritional relevance for humanity. The plant cuticle is the external lipid layer covering the nonwoody aerial organs of land plants, and it is the first contact between fruits and the environment. It has been hypothesized that the cuticle plays a role in the development, ripening, quality, resistance to pathogen attack and postharvest shelf life of fleshy fruits. The cuticle's structure and composition change in response to the fruit's developmental stage, fruit physiology and different postharvest treatments. This review summarizes current information on the physiology and molecular mechanism of cuticle biosynthesis and composition changes during the development, ripening and postharvest stages of fleshy fruits. A discussion and analysis of studies regarding the relationship between cuticle composition, water loss reduction and maintaining fleshy fruits' postharvest quality are presented. An overview of the molecular mechanism of cuticle biosynthesis and efforts to elucidate it in fleshy fruits is included. Enhancing our knowledge about cuticle biosynthesis mechanisms and identifying specific transcripts, proteins and lipids related to quality traits in fleshy fruits could contribute to the design of biotechnological strategies to improve the quality and postharvest shelf life of these important fruit crops. [ABSTRACT FROM AUTHOR]

In the Ethiopian highlands, clearance of Afromontane dry forest and conversion to crop and grazing land lead to land degradation and loss of soil organic matter (SOM). Eucalyptus is often grown on degraded soils, and this results in the partial recovery of soil carbon stocks. The aim of this work was to assess the biological sources of SOM in this land-use sequence. In top-soils (0–10 cm) of four land-use systems, namely remnant natural forest, eucalyptus plantation, cropland, and grazing land, in the Ethiopian highlands, the origin of SOM was investigated. For this, a sequential extraction method was used, involving a solvent extraction, base hydrolysis, and a subsequent CuO oxidation. In these extracts, biomarkers (molecular proxies) were identified to characterize the SOM of the soil of the four land-uses. Putative lipid monomers of leaf, root, and microbial degradation products suggest that root inputs and microbial inputs dominate in SOM of all the land-uses, except grazing land. The ratios of syringyls, vanillyls, and cinnamyls showed that non-woody angiosperm plants were the predominant source for lignin in eucalyptus, cropland, and grazing land soil. In the soils of the natural forest, lignin originates from both woody angiosperms and woody gymnosperms. Our study shows the importance of root and microbial inputs in the formation of SOM, but also that, in the natural forest, legacies of previous forest cover are present. [ABSTRACT FROM AUTHOR]

Background and objectives: Micromorphological and elemental information on pulse cotyledon topographies is an essential aspect in identifying grain physiological and structural interactions at cotyledon‐seed coat (abaxial) and cotyledon–cotyledon (adaxial) interfaces, which can influence grain quality and processing behavior in food industry applications. The objective of this study was to characterize the topographies of the abaxial and adaxial surfaces of two cultivars each of faba bean, chickpea, field pea, and lentil via micromorphological and elemental analyses. Findings: Scanning electron microscopic imaging of the abaxial and adaxial cotyledon surfaces showed gentle undulation of either tabular‐ or polygonal‐shaped epidermal cells that were projected as flat or rugose. They were covered by epicuticular wax films, which had smooth, striate, or granular micromorphology. Elemental results confirmed the presence of C, O, K, Ca, and Mg as the key elements on both cotyledon abaxial and adaxial surfaces. Conclusions: Abaxial and adaxial cotyledon topographies of all the cultivars confirmed varietal differences associated with their micromorphological characteristics. It can be concluded that both abaxial and adaxial surfaces are cemented by a complex cuticle layer of similar elemental composition with a few significant (p <.05) variations. The ease of milling/pearling of pulses could be directly associated with the species, cultivar, architecture, micromorphology, and elemental composition of the pulse cotyledon abaxial and adaxial surfaces. Significance and novelty: These findings will not only assist in interpreting the differences observed in the processing behavior of different pulse species and cultivars but will also enhance our understanding of the process design, targeting pearling of pulses for value‐added food applications. [ABSTRACT FROM AUTHOR]

Cuticles are specialized cell wall structures that form at the surface of terrestrial plant organs. They are largely comprised lipidic compounds and are deposited in the apoplast, external to the polysaccharide-rich primary wall, creating a barrier to diffusion of water and solutes, as well as to environmental factors. The predominant cuticle component is cutin, a polyester that is assembled as a complex matrix, within and on the surface of which aliphatic and aromatic wax molecules accumulate, further modifying its properties. To reach the point of cuticle assembly the different acyl lipid-containing components are first exported from the cell across the plasma membrane and then traffic across the polysaccharide wall. The export of cutin precursors and waxes from the cell is known to involve plasma membrane-localized ATP-binding cassette (ABC) transporters; however, other secretion mechanisms may also contribute. Indeed, extracellular vesiculo-tubular structures have recently been reported in Arabidopsis thaliana (Arabidopsis) to be associated with the deposition of suberin, a polyester that is structurally closely related to cutin. Intriguingly, similar membranous structures have been observed in leaves and petals of Arabidopsis, although in lower numbers, but no close association with cutin formation has been identified. The possibility of multiple export mechanisms for cuticular components acting in parallel will be discussed, together with proposals for how cuticle precursors may traverse the polysaccharide cell wall before their assimilation into the cuticle macromolecular architecture. [ABSTRACT FROM AUTHOR]

The cuticle covers almost all plant organs as the outermost layer and serves as a transpiration barrier, sunscreen, and first line of defense against pathogens. Waxes, fatty acids, and aromatic components build chemically and structurally diverse layers with different functionality. So far, electron microscopy has elucidated structure, while isolation, extraction, and analysis procedures have revealed chemistry. With this method paper, we close the missing link by demonstrating how Raman microscopy gives detailed information about chemistry and structure of the native cuticle on the microscale. We introduce an optimized experimental workflow, covering the whole process of sample preparation, Raman imaging experiment, data analysis, and interpretation and show the versatility of the approach on cuticles of a spruce needle, a tomato peel, and an Arabidopsis stem. We include laser polarization experiments to deduce the orientation of molecules and multivariate data analysis to separate cuticle layers and verify their molecular composition. Based on the three investigated cuticles, we discuss the chemical and structural diversity and validate our findings by comparing models based on our spectroscopic data with the current view of the cuticle. We amend the model by adding the distribution of cinnamic acids and flavonoids within the cuticle layers and their transition to the epidermal layer. Raman imaging proves as a non-destructive and fast approach to assess the chemical and structural variability in space and time. It might become a valuable tool to tackle knowledge gaps in plant cuticle research. [ABSTRACT FROM AUTHOR]

Terrestrialization of vascular plants, i.e., Angiosperm, is associated with the development of cuticular barriers that prevent biotic and abiotic stresses and support plant growth and development. To fulfill these multiple functions, cuticles have developed a unique supramolecular and dynamic assembly of molecules and macromolecules. Plant cuticles are not only an assembly of lipid compounds, i.e., waxes and cutin polyester, as generally presented in the literature, but also of polysaccharides and phenolic compounds, each fulfilling a role dependent on the presence of the others. This mini-review is focused on recent developments and hypotheses on cuticle architecture–function relationships through the prism of non-lipid components, i.e., cuticle-embedded polysaccharides and polyester-bound phenolics. [ABSTRACT FROM AUTHOR]