Cuticle Permeability Research Articles

Wheat Topoisomerase VI Positively Regulates the Biosynthesis of Cuticular Wax and Cutin.

Lipophilic cuticles mainly composed of wax mixtures and cutin matrices seal the plant epidermis and control plant development and environmental adaptation. Although cuticle-associated traits have been selected in the breeding of agronomically important cereal bread wheat, the biosynthesis of wheat cuticular wax and cutin remains poorly understood. Herein, wheat topoisomerase VI was characterized as an essential activator of cuticular wax and cutin biosynthesis. Knock-down of wheat TaTOP6A, TaTOP6B, TaRHL1, or TaBIN4 gene encoding component of topoisomerase VI resulted in decreased loads of leaf cuticular wax and cutin, as well as increased leaf cuticle permeability. Moreover, TaCYP86A2 was identified as a key component of the wheat cutin biosynthetic machinery. Reduction of wheat TaCYP86A2 expression led to decreased cutin accumulation and enhanced cuticle permeability. In addition, TaTOP6A, TaTOP6B, TaRHL1, or TaBIN4 was shown to enrich at the promoter regions of the wax biosynthesis gene TaKCS1 and the cutin biosynthesis gene TaCYP86A2. Importantly, chromatin at TaKCS1 and TaCYP86A2 promoters is marked by high nucleosome occupancy and low histone acetylation in TaTOP6A-, TaTOP6B-, TaRHL1-, or TaBIN4-silenced wheat leaves. These results collectively support that wheat topoisomerase VI positively regulates the biosynthesis of cuticular wax and cutin probably via maintaining a permissive chromatin state at TaKCS1 and TaCYP86A2 genes.

Arabidopsis DREB26/ERF12 and its close relatives regulate cuticular wax biosynthesis under drought stress condition.

Land plants have evolved a hydrophobic cuticle on the surface of aerial organs as an adaptation to ensure survival in terrestrial environments. Cuticle is mainly composed of lipids, namely cutin and intracuticular wax, with epicuticular wax deposited on plant surface. The composition and permeability of cuticle have a large influence on its ability to protect plants against drought stress. However, the regulatory mechanisms underlying cuticular wax biosynthesis in response to drought stress have not been fully elucidated. Here, we identified three AP2/ERF transcription factors (DREB26/ERF12, ERF13 and ERF14) involved in the regulation of water permeability of the plant surface. Transmission electron microscopy revealed thicker cuticle on the leaves of DREB26-overexpressing (DREB26OX) plants, and thinner cuticle on the leaves of transgenic plants expressing SRDX repression domain-fused DREB26 (DREB26SR). Genes involved in cuticular wax formation were upregulated in DREB26OX and downregulated in DREB26SR. The levels of very-long chain (VLC) alkanes, which are a major wax component, increased in DREB26OX leaves and decreased in DREB26SR leaves. Under dehydration stress, water loss was reduced in DREB26OX and increased in DREB26SR. The erf12/13/14 triple mutant showed delayed growth, decreased leaf water content, and reduced drought-inducible VLC alkane accumulation. Taken together, our results indicate that the DREB26/ERF12 and its closed family members, ERF13 and ERF14, play an important role in cuticular wax biosynthesis in response to drought stress. The complex transcriptional cascade involved in the regulation of cuticular wax biosynthesis under drought stress conditions is discussed.

Wheat Transcription Factor TaMYB60 Modulates Cuticular Wax Biosynthesis by Activating TaFATB and TaCER1 Expression.

Cuticular wax mixtures cover the epidermis of land plants and shield plant tissues from abiotic and biotic stresses. Although cuticular wax-associated traits are employed to improve the production of bread wheat, regulatory mechanisms underlying wheat cuticular wax biosynthesis remain poorly understood. In this research, partially redundant transcription factors TaMYB60-1 and TaMYB60-2 were identified as positive regulators of wheat cuticular wax biosynthesis. Knock-down of wheat TaMYB60-1 and TaMYB60-2 genes by virus-induced gene silencing resulted in attenuated wax accumulation and enhanced cuticle permeability. The roles of wheat fatty acyl-ACP thioesterase genes TaFATB1 and TaFATB2 in cuticular wax biosynthesis were characterized. Silencing wheat TaFATB1 and TaFATB2 genes led to reduced wax accumulation and increased cuticle permeability, suggesting that TaFATB1 and TaFATB2 genes positively contribute to wheat cuticular wax biosynthesis. Importantly, transcription factors TaMYB60-1 and TaMYB60-2 exhibit transcriptional activation ability and could stimulate the expression of wax biosynthesis genes TaFATB1, TaFATB2, and ECERIFERUM 1 (TaCER1). These findings support that transcription factor TaMYB60 positively regulates wheat cuticular wax biosynthesis probably by activating transcription of TaFATB1, TaFATB2, and TaCER1 genes.

SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 13 (SLP13) together with SPL9 redundantly regulates wax biosynthesis under drought stress.

Wax biosynthesis is closely controlled by many regulators under different environmental conditions. We have previously shown that the module miR156-SQUAMOSA PROMOTER BINDING PROTEIN-LIKE9 (SPL9)-DEWAX is involved in the diurnal regulation of wax production; however, it was not determined whether other SPLs are also involved in wax synthesis. Here, we report that SPL13 also regulates drought-induced wax production, by directly and indirectly affecting the expression of the two wax biosynthesis genes ECERIFERUM1 (CER1) and CER4, respectively. In addition, we show that SPL13 together with SPL9 redundantly regulates wax accumulation under both normal and drought stress conditions, and that simultaneous mutation of both genes additively increases cuticle permeability and decreases drought tolerance. However, in contrast to SPL9, SPL13 does not seem to participate in the DEWAX-mediated diurnal regulation of wax production.

Identification of long-chain acyl-CoA synthetase gene family reveals that SlLACS1 is essential for cuticular wax biosynthesis in tomato

Long-chain acyl-CoA synthetases (LACSs), belonging to the acyl-activating enzyme superfamily, play crucial roles in lipid biosynthesis and fatty acid catabolism. Here, we identified 11 LACS genes in the tomato reference genome, and these genes were clustered into six subfamilies. Gene structure and conserved motif analyses indicated that LACSs from the same subfamily shared conserved gene and protein structures. Expression analysis revealed that SlLACS1 was highly expressed in the outer epidermis of tomato fruits and leaves. Subcellular localization assay results showed that SlLACS1 was located in the endoplasmic reticulum. Compared with wild-type plants, the wax content on leaves and fruits decreased by 22.5–34.2 % in SlLACS1 knockout lines, confirming that SlLACS1 was involved in wax biosynthesis in both leaves and fruits. Water loss, chlorophyll extraction, water-deficit, and toluidine blue assays suggested that cuticle permeability was elevated in SlLACS1 knockout lines, resulting in reduction in both drought stress resistance and fruit shelf-life. Overall, our analysis of the LACSs in tomato, coupled with investigations of SlLACS1 function, yielded a deeper understanding of the evolutionary patterns of LACS members and revealed the involvement of SlLACS1 in wax accumulation contribute to drought resistance and extended fruit shelf-life in tomato.

3-ketoacyl-CoA synthase 19 contributes to the biosynthesis of seed lipids and cuticular wax in Arabidopsis and abiotic stress tolerance.

Very-long-chain fatty acids (VLCFAs) are essential precursors for plant membrane lipids, cuticular waxes, suberin, and storage oils. Integral to the fatty acid elongase (FAE) complex, 3-ketoacyl-CoA synthases (KCSs) function as crucial enzymes in the VLCFA pathway, determining the chain length of VLCFA. This study explores the in-planta role of the KCS19 gene. KCS19 is predominantly expressed in leaves and stem epidermis, sepals, styles, early silique walls, beaks, pedicels, and mature embryos. Localized in the endoplasmic reticulum, KCS19 interacts with other FAE proteins. kcs19 knockout mutants displayed reduced total wax and wax crystals, particularly alkanes, while KCS19 overexpression increased these components and wax crystals. Moreover, the cuticle permeability was higher for the kcs19 mutants compared to the wild type, rendering them more susceptible to drought and salt stress, whereas KCS19 overexpression enhanced drought and salt tolerance. Disrupting KCS19 increased C18 species and decreased C20 and longer species in seed fatty acids, indicating its role in elongating C18 to C20 VLCFAs, potentially up to C24 for seed storage lipids. Collectively, KCS19-mediated VLCFA synthesis is required for cuticular wax biosynthesis and seed storage lipids, impacting plant responses to abiotic stress.

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

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

Cuticle properties, wax composition, and crystal morphology of Hami melon cultivars (Cucumis melo L.) with differential resistance to fruit softening

Cuticle wax chemicals are cultivar-dependent and contribute to storage quality. Few research reported on wax analysis between melting flesh-type (MF; ‘Jinhuami 25’) and nonmelting flesh-type (NMF; ‘Xizhoumi 17’ and ‘Chougua’) Hami melons. Chemicals and crystal structures of Hami melon cuticular wax, cell wall metabolism related to fruit melting, and fruit physiology were analyzed to observe wax functions. Results showed that Hami melon cuticle wax predominantly consists of esters, alkanes, alcohols, aldehydes, and terpenoids. MF-type has a lower alkane/terpenoid ratio, concomitant to its higher weight loss and cuticle permeability. Micromorphology of wax crystals appears as numerous platelets with irregular crystals, and the transformation of wax structure in NMF Hami melon is delayed. Waxy components affect cell wall metabolism and physiological quality, which results in the pulp texture difference between MF-type and NMF-type during storage. Results provide a reference for the regulation of wax synthesis in both types of melons.

Diatomaceous earth as insecticide: physiological and morphological evidence of its underlying mechanism.

Wheat grain containers or silos can be perfect habitats for insects, which generate large economic losses to grain production. Natural alternatives to synthetic insecticides have grown in popularity because of health, economic and ecological issues. Diatomaceous earth is a natural compound that has an insecticide effect by enhancing an insect's dehydration with no toxicity on mammals including humans. The aim of this study is to confirm the effect of diatomaceous earth as an insecticide for the wheat grain pest, the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae) and demonstrate its underlying mechanisms as an insecticide by open-flow respirometry and scanning electron microscopy. Survival bioassays of T. castaneum revealed a dose-dependent insecticide effect of diatomaceous earth. Gravimetric measurements showed that 2 days exposure to diatomaceous earth produces a significant increase of mass loss. Open-flow respirometry measurements showed an increase of total water emission rate on insects due to an increase of both, respiratory and cuticular water loss. Our study revealed that diatomaceous earth produces an increase of insect's cuticle permeability, which is responsible for elevated cuticular water loss. Scanning electron microscopy images provided visual evidence of the lipid absorbent properties of diatomaceous earth particles, and showed a tendency for higher, although not significant, damaged area of the cuticle's surface from diatomaceous earth treated insects compared to control ones. With state-of-the art techniques like open-flow respirometry and scanning electron microscopy, we demonstrated the underlying mechanism of diatomaceous earth as an insecticide and provided new cues for understanding the properties of the cuticle and its ecological importance. © 2024 Society of Chemical Industry.

Do Cuticular Gaps Make It Possible to Study the Composition of the Cell Walls in the Glands of Drosophyllum lusitanicum?

Carnivorous plants can survive in poor habitats because they have the ability to attract, capture, and digest prey and absorb animal nutrients using modified organs that are equipped with glands. These glands have terminal cells with permeable cuticles. Cuticular discontinuities allow both secretion and endocytosis. In Drosophyllum lusitanicum, these emergences have glandular cells with cuticular discontinuities in the form of cuticular gaps. In this study, we determined whether these specific cuticular discontinuities were permeable enough to antibodies to show the occurrence of the cell wall polymers in the glands. Scanning transmission electron microscopy was used to show the structure of the cuticle. Fluorescence microscopy revealed the localization of the carbohydrate epitopes that are associated with the major cell wall polysaccharides and glycoproteins. We showed that Drosophyllum leaf epidermal cells have a continuous and well-developed cuticle, which helps the plant inhibit water loss and live in a dry environment. The cuticular gaps only partially allow us to study the composition of cell walls in the glands of Drosophyllum. We recoded arabinogalactan proteins, some homogalacturonans, and hemicelluloses. However, antibody penetration was only limited to the cell wall surface. The localization of the wall components in the cell wall ingrowths was missing. The use of enzymatic digestion improves the labeling of hemicelluloses in Drosophyllum glands.

A cuticular protein, BgCPLCP1, contributes to insecticide resistance by thickening the cockroach endocuticle

Overuse of insecticides has led to severe environmental problems. Insect cuticle, which consists mainly of chitin, proteins and a thin outer lipid layer, serves multiple functions. Its prominent role is as a physical barrier that impedes the penetration of xenobiotics, including insecticides. Blattella germanica (L.) is a major worldwide indoor pest that causes allergic disease and asthma. Extensive use of pyrethroid insecticides, including β-cypermethrin, has selected for the rapid and independent evolution of resistance in cockroach populations on a global scale. We demonstrated that BgCPLCP1, the first CPLCP (cuticular proteins of low complexity with a highly repetitive proline-rich region) family cuticular protein in order Blattodea, contributes to insecticide penetration resistance. Silencing BgCPLCP1 resulted in 85.0 %–85.7 % and 81.0 %–82.0 % thinner cuticle (and especially thinner endocuticle) in the insecticide-susceptible (S) and β-cypermethrin-resistant (R) strains, respectively. The thinner and more permeable cuticles resulted in 14.4 % and 20.0 % lower survival of β-cypermethrin-treated S- and R-strain cockroaches, respectively. This study advances our understanding of cuticular penetration resistance in insects and opens opportunities for the development of new efficiently and environmentally friendly insecticides targeting the CPLCP family of cuticular proteins.

LTP2 hypomorphs show genotype-by-environment interaction in early seedling traits in Arabidopsis thaliana.

Isogenic individuals can display seemingly stochastic phenotypic differences, limiting the accuracy of genotype-to-phenotype predictions. The extent of this phenotypic variation depends in part on genetic background, raising questions about the genes involved in controlling stochastic phenotypic variation. Focusing on early seedling traits in Arabidopsis thaliana, we found that hypomorphs of the cuticle-related gene LIPID TRANSFER PROTEIN 2 (LTP2) greatly increased variation in seedling phenotypes, including hypocotyl length, gravitropism and cuticle permeability. Many ltp2 hypocotyls were significantly shorter than wild-type hypocotyls while others resembled the wild-type. Differences in epidermal properties and gene expression between ltp2 seedlings with long and short hypocotyls suggest a loss of cuticle integrity as the primary determinant of the observed phenotypic variation. We identified environmental conditions that reveal or mask the increased variation in ltp2 hypomorphs and found that increased expression of its closest paralog LTP1 is necessary for ltp2 phenotypes. Our results illustrate how decreased expression of a single gene can generate starkly increased phenotypic variation in isogenic individuals in response to an environmental challenge.

Fine-tuning the activities of β-KETOACYL-COA SYNTHASE 3 (KCS3) and KCS12 in Arabidopsis is essential for maintaining cuticle integrity.

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.

Adsorption and permeability of heavy metals (Fe, Cu, Pb, Zn, Cr, and Cd) onto the adaxial cuticle of Ficus elastica leaf

Air pollution represents a serious worldwide problem as atmospheric deposition of pollutants widely affects human and wildlife health. Heavy metals are significant pollutants with high toxicity, persistence, and bioaccumulation. In an environment polluted by industry and transportation, deposits of heavy metals on the leaves and their subsequent uptake by the plant occur. The leaf cuticle is the most important barrier between plant tissue and the atmosphere. To understand the processes of adsorption of heavy metals and their diffusion into the leaf, adaxial cuticles of Ficus elastica were enzymatically isolated and used in batch adsorption experiments and permeability measurements using diffusion chambers. A demonstration experiment with a whole leaf was also carried out, which pointed to the real uptake of metals by the leaf. Cuticles were subjected to characterization by methods of elemental analysis, Fourier transform infrared spectroscopy, and scanning electron microscopy. In the adsorption experiment, significant differences in the maximum adsorption capacity (Cd>Pb>Cr>Fe>Cu>Zn), and mechanism of adsorption (Langmuir model for Fe and Cr, Freundlich model for Zn, Cu, Pb, and Cd) were found. The permeability of intact adaxial cuticle for studied heavy metals did not differ significantly. The extent of foliar uptake differed between metals, with the relatively highest uptake of Fe and Zn. The transport of metals in the leaf was also observed. It seems that adsorption to the leaf surface more than the permeability of the cuticle is a limiting factor for foliar uptake of heavy metals from atmospheric deposits.

OsFAR1 is involved in primary fatty alcohol biosynthesis and promotes drought tolerance in rice.

OsFAR1 encodes a fatty acyl-CoA reductase involved in biosynthesis of primary alcohols and plays an important role in drought stress response in rice. Cuticular waxes cover the outermost surface of terrestrial plants and contribute to inhibiting nonstomatal water loss and improving plant drought resistance. Primary alcohols are the most abundant components in the leaf cuticular waxes of rice (Oryza sativa), but the biosynthesis and regulation of primary alcohol remain largely unknown in rice. Here, we identified and characterized an OsFAR1 gene belonging to the fatty acyl-CoA reductases (FARs) via a homology-based approach in rice. OsFAR1 was activated by abiotic stresses and abscisic acid, resulting in increased production of primary alcohol in rice. Heterologous expression of OsFAR1 enhanced the amounts of C22:0 and C24:0 primary alcohols in yeast (Saccharomyces cerevisiae) and C24:0 to C32:0 primary alcohols in Arabidopsis. Similarly, OsFAR1 overexpression significantly increased the content of C24:0 to C30:0 primary alcohols on rice leaves. Finally, OsFAR1 overexpression lines exhibited reduced cuticle permeability and enhanced drought tolerance in rice and Arabidopsis. Taken together, our results demonstrate that OsFAR1 is involved in rice primary alcohol biosynthesis and plays an important role in responding to drought and other environmental stresses.

Transcription Factor TaMYB30 Activates Wheat Wax Biosynthesis.

The waxy cuticle covers a plant's aerial surface and contributes to environmental adaptation in land plants. Although past decades have seen great advances in understanding wax biosynthesis in model plants, the mechanisms underlying wax biosynthesis in crop plants such as bread wheat remain to be elucidated. In this study, wheat MYB transcription factor TaMYB30 was identified as a transcriptional activator positively regulating wheat wax biosynthesis. The knockdown of TaMYB30 expression using virus-induced gene silencing led to attenuated wax accumulation, increased water loss rates, and enhanced chlorophyll leaching. Furthermore, TaKCS1 and TaECR were isolated as essential components of wax biosynthetic machinery in bread wheat. In addition, silencing TaKCS1 and TaECR resulted in compromised wax biosynthesis and potentiated cuticle permeability. Importantly, we showed that TaMYB30 could directly bind to the promoter regions of TaKCS1 and TaECR genes by recognizing the MBS and Motif 1 cis-elements, and activate their expressions. These results collectively demonstrated that TaMYB30 positively regulates wheat wax biosynthesis presumably via the transcriptional activation of TaKCS1 and TaECR.

Larva morphology of shore flies <i>Ephydra riparia</i> and <i>Paracoenia fumosa</i> (Diptera: Ephydridae) and adaptation of Diptera to increased salinity

Larvae of many shore fly species (family Ephydridae) are adapted to living in water with high or extremely high salinity. Little is known about the morphological and physiological foundations of such adaptation. We described the details of the morphology of third-instar larvae of two shore flies: Ephydra riparia and Paracoenia fumosa and presented the scanning electron microscopy (SEM) images. For the first time, by silver-staining and SEM, we proved that the larvae of both studied species had anal organs (AO) – specialized structures that serve an osmoregulatory function and are responsible for the transport of ions from the environment to the larval hemolymph (but not in the opposite direction). We compared the larvae morphology of the studied species with some other shore fly species from the genera Ephydra, Paracoenia, Hydrellia, and Coenia, as well as with larvae of the model species Drosophila melanogaster (family Drosophilidae). Special attention was paid to the morphology of AO, which contribute to the adaptation of larvae to increased salinity. Extremely halophilic species either do not have AO, or they are poorly developed, while moderately halophilic shore flies have more developed features connected with the permeability of the AO cuticle and active ion transport. These features are most developed in freshwater shore flies. AO activity can vary due to the shape and area of the AO, the smoothness or wrinkling of the cuticle, and the presence of nanoscale pits on it. Described variability of AO structure is probably adaptive since, at high salinity, both the permeability of the AO cuticle and the active transport of ions from the environment to the hemolymph become less beneficial or even harmful.

Apple SUMO E3 ligase MdSIZ1 regulates cuticular wax biosynthesis by SUMOylating transcription factor MdMYB30.

A key function of SUMOylation is the coordinated modification of numerous proteins to optimize plant growth and resistance to environmental stress. Plant cuticular wax is deposited on the surface of primary plant organs to form a barrier that provides protection against changes in terrestrial environments. Many recent studies have examined cuticular wax biosynthetic pathways and regulation. However, whether SUMOylation is involved in the regulation of cuticle wax deposition at the posttranslational level remains unclear. Here, we demonstrate that a small ubiquitin-like modifier (SUMO) E3 ligase, SAP AND MIZ1 DOMAIN CONTAINING LIGASE1 (MdSIZ1), regulates wax accumulation and cuticle permeability in apple (Malus domestica Borkh), SUMO E2 CONJUGATING ENZYME 1(MdSCE1) physically interacts with MdMYB30, a transcription factor involved in the regulation of cuticle wax accumulation. MdSIZ1 mediates the SUMOylation and accumulation of MdMYB30 by inhibiting its degradation through the 26S proteasome pathway. Furthermore, MdMYB30 directly binds to the β-KETOACYL-COA SYNTHASE 1 (MdKCS1) promoter to activate its expression and promote wax biosynthesis. These findings indicate that the MdSIZ1-MdMYB30-MdKCS1 module positively regulates cuticular wax biosynthesis in apples. Overall, the findings of our study provide insights into the regulation pathways involved in cuticular wax biosynthesis.

CsCER6 and CsCER7 Influence Fruit Glossiness by Regulating Fruit Cuticular Wax Accumulation in Cucumber.

Fruit glossiness is an important external fruit quality trait that greatly affects the marketability of fresh cucumber (Cucumis sativus) fruits. A few reports have suggested that the extent of cuticular wax loading influences the glossiness of the fruit surface. In the present study, we tested the wax contents of two inbred cucumber lines, comparing a line with waxy fruit (3401) and a line with glossy fruit (3413). Wax content analysis and dewaxing analysis demonstrate that fruit cuticular wax loads negatively correlate with fruit glossiness in cucumber. Identifying genes that were differentially expressed in fruit pericarps between 3401 and 3413 and genes induced by abscisic acid suggested that the wax biosynthesis gene CsCER6 (Cucumis sativus ECERIFERUM 6) and the regulatory gene CsCER7 may affect wax accumulation on cucumber fruit. Expression analysis via RT-qPCR, GUS-staining, and in situ hybridization revealed that CsCER6 and CsCER7 are abundantly expressed in the epidermis cells in cucumber fruits. Furthermore, the overexpression and RNAi lines of CsCER6 and CsCER7 showed dramatic effects on fruit cuticular wax contents, fruit glossiness, and cuticle permeability. Our results suggest that CsCER6 and CsCER7 positively regulate fruit cuticular wax accumulation and negatively influence fruit glossiness.

Differences in leaf cuticular wax induced by whole-genome duplication in autotetraploid sour jujube

Drought-resistant plants exhibit strong water retention capability. In this regard, the autotetraploid sour jujube leaves exhibit better water retention than diploid leaves. Morphological comparisons and physiological comparisons of diploid and autotetraploid leaves showed that the autotetraploid leaves had thicker leaf cuticles and more leaf wax accumulation than the diploid leaves, which could reduce cuticle permeability and improve the drought tolerance of leaves. In this study, the cuticular wax crystalloids on the adaxial and abaxial sides of young and mature jujube leaves were observed in the two ploidy types, and unique cuticular wax crystalloids covering a large area of the cuticle on autotetraploid sour jujube leaves may provide an advantage in reducing leaf non-stomata transpiration and improving plant drought tolerance. Based on the transcriptome, 115 differentially expressed genes between diploids and autotetraploids were further analyzed and found to be involved in the accumulation of cuticular wax components, including terpenoids, fatty acids, and lipids, as well as ABC transporter and wax biosynthetic process. Finally, 14 genes differentially expressed between glossy autotetraploid leaves and nonglossy diploid leaves, such as LOC107414787, LOC107411574 and LOC107413721, were screened as candidate genes by qRT-PCR analysis. This findings provided insights into how polyploidization improved drought tolerance.