Non-stomatal Water Loss Research Articles

BrWax1 function in wax biosynthesis was verified by allelic mutations in Chinese cabbage

The primary function of cuticular wax is to prevent non-stomatal water loss, while the wax deficiency conferring a glossy appearance on plants is conductive to crop commodity qualities. We previously mapped a wax-deficient gene BrWax1 in a natural mutant ‘08A235-2’ by SSR marker, and predicted Bra013809 (BrWax1) as the candidate gene in Chinese cabbage (Brassica rapa L. ssp. pekinensis). In order to verifying the function of BrWax1, two allelic wax-crystal deficiency mutants wdm4 and wdm8 from an EMS mutagenesis population of Chinese cabbage were screened. wdm4 was applied to identify the mutant gene, with the finding that BraA01g015290.3C, the homologous of AT4G24510 (AtCER2), was the candidate gene. Bra013809 in Brara_Chiifu_V1.5 and BraA01g015290.3C in Brara_Chiifu_V3.0 were annotated as the same gene in Brassicaceae Database. A SNP (C to T) on BrWax1 in wdm4 resulted in a stopgain, and a SNP (G to A) on BrWax1 in wdm8 led to the deletion of 38 bases which caused a stopgain at the 190th amino acid. These results finally verified the function of BrWax1 in the wax biosynthesis. Our findings emphasize the crucial role of the BrWax1 gene in cuticular wax biosynthesis in Chinese cabbage and suggest its potential as a valuable genetic resource for breeding for glossy appearance.

Micromorphological and Chemical Characterization of Drimys winteri Leaf Surfaces: The Secondary Alcohols Forming Epicuticular Wax Crystals Are Accompanied by Alkanediol, Alkanetriol and Ketol Derivatives.

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.

Revealing the crucial role of cuticular wax components in postharvest water loss in passion fruit (Passiflora edulis Sims.)

Passion fruit (Passiflora edulis Sims.) is highly prone to water loss and shrinkage after harvesting. The fruit cuticle serves as a crucial barrier to prevent non-stomatal water loss. However, the impact of cuticle on water loss in passion fruit remains unclear. Therefore, the present study primarily focuses on the influence of the cuticle on postharvest water loss of passion fruit, specifically emphasizing the changes in cuticular wax components. Our results showed that both the thickness and content of cutin decreased during storage. The degradation of wax components exhibited a similar trend to fruit water loss. At the end of the storage, n-triacontane content was stored at 25 °C retained 2.8% of that at 7 °C. Follwed by stearic acid, palmitate, and so on, with amounts at 25 °C are 9.3% and 13.8% at 7 °C, respectively. Interestingly, the reduction of amides, erucamide and hexadecanamide lead to water loss in fruit. So far as we know, this is the first report demonstrating that cuticle amides influence water loss in fruit. This study provides new insights into the role of cuticle in the water loss of passion fruit.

AtMYB41 acts as a dual-function transcription factor that regulates the formation of lipids in an organ- and development-dependent manner.

The plant cuticle controls non-stomatal water loss and can serve as a barrier against biotic agents, whereas the heteropolymer suberin and its associated waxes are deposited constitutively at specific cell wall locations. While several transcription factors controlling cuticle formation have been identified, those involved in the transcriptional regulation of suberin biosynthesis remain poorly characterized. The major goal of this study was to further analyse the function of the R2R3-Myeloblastosis (MYB) transcription factor AtMYB41 in formation of the cuticle, suberin, and suberin-associated waxes throughout plant development. For functional analysis, the organ-specific expression pattern of AtMYB41 was analysed and Atmyb41ge alleles were generated using the CRISPR/Cas9 system. These were investigated for root growth and water permeability upon stress. In addition, the fatty acid, wax, cutin, and suberin monomer composition of different organs was evaluated by gas chromatography. The characterization of Atmyb41ge mutants revealed that AtMYB41 negatively regulates the production of cuticular lipids and fatty acid biosynthesis in leaves and seeds, respectively. Remarkably, biochemical analyses indicate that AtMYB41 also positively regulates the formation of cuticular waxes in stems of Arabidopsis thaliana. Overall, these results suggest that the AtMYB41 acts as a negative regulator of cuticle and fatty acid biosynthesis in leaves and seeds, respectively, but also as a positive regulator of wax production in A. thaliana stems.

Transcription factors and plant hormones mediate wax metabolism in response to drought stress.

Plants have, throughout evolution, developed a hydrophobic cuticle to protect them from various stresses in the terrestrial environment. The cuticle layer is mainly composed of cutin and cuticular wax, a mixture of very-long-chain fatty acids and their derivatives. With the progress of transcriptome sequencing and other research methods, the key enzymes, transporters and regulatory factors in wax synthesis and metabolism have been gradually identified, especially the study on the regulation of wax metabolism by transcription factors and others in response to plant stress has become a hot topic. Drought is a major abiotic stress that limits plant growth and crop productivity. Plant epidermal wax prevents non-stomatal water loss and improves water use efficiency to adapt to arid environments. In this study, the ways of wax synthesis, transport, metabolism and regulation at different levels are reviewed. At the same time, the regulation of wax by different transcription factors and plant hormones in response to drought is elaborated, and key research questions and important directions for future solutions are proposed to enhance the potential application of epidermal wax in agriculture and the environment.

β-diketone accumulation in response to drought stress is weakened in modern bread wheat varieties (Triticum aestivum L.).

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

Phenotypic Diversity in Leaf Cuticular Waxes in Brassica carinata Accessions.

Brassica carinata has received considerable attention as a renewable biofuel crop for semi-arid zones due to its high oil content and polyunsaturated fatty acids contents. It is important to develop new drought-resistant cultivars of B. carinata production to expand its areas into more arid regions. The accumulation of leaf cuticular wax on plant surfaces is one mechanism that reduces non-stomatal water loss, thus increasing drought resistance in plants. To explore phenotypic variations in cuticular wax in B. carinata, leaf waxes were extracted and quantified from a diversity panel consisting of 315 accessions. The results indicate that the accessions have a wide range of total leaf wax content (289-1356 µg dm-2), wax classes, and their components. The C29 and C31 homologues of alkanes, C29 ketone homologue, C29 secondary alcohol, and C30 aldehyde were the most abundant leaf waxes extracted from B. carinata accessions. The high heritability values of these waxes point to the positive selection for high wax content during early generations of future B. carinata breeding programs. Positive correlation coefficients, combined with the effects of these waxes on leaf wax content accumulation, suggest that modifying specific wax content could increase the total wax content and enhance cuticle composition. The identified leaf wax content and compositions in B. carinata will lead to the future discovery of wax biosynthetic pathways, the dissection of its genetic regulatory networks, the identification of candidate genes controlling production of these waxes, and thus, develop and release new B. carinata drought-tolerant cultivars.

A 3-Ketoacyl-CoA Synthase 10 (KCS10) Homologue from Alfalfa Enhances Drought Tolerance by Regulating Cuticular Wax Biosynthesis.

Cuticular wax, forming the first line of defense against adverse environmental stresses, comprises very long-chain fatty acids (VLCFAs) and their derivatives. 3-Ketoacyl-CoA synthase (KCS) is a rate-limiting enzyme for VLCFA biosynthesis. In this study, we isolated KCS10, a KCS gene from alfalfa, and analyzed the effect of gene expression on wax production and drought stress in transgenic plants. MsKCS10 overexpression increased compact platelet-like crystal deposition and promoted primary alcohol biosynthesis through acyl reduction pathways in alfalfa leaves. Overexpression of MsKCS10 induced the formation of coiled-rodlet-like crystals and increased n-alkane content through decarbonylation pathways in tobacco and tomato fruits. Overexpression of MsKCS10 enhanced drought tolerance by limiting nonstomatal water loss, improving photosynthesis, and maintaining osmotic potential under drought stress in transgenic tobacco. In summary, MsKCS10 plays an important role in wax biosynthesis, wax crystal morphology, and drought tolerance, although the mechanisms are different among the plant species. MsKCS10 can be targeted in future breeding programs to improve drought tolerance in plants.

Distinct epiphyte responses to drought in tropical mountain cloud forests

AbstractEpiphytes are often considered to be significantly impacted by precipitation changes because their lack of direct access to soil water. However, few in situ studies have examined how reduction in water availability may affect epiphyte growth. Using a unique stemflow collection and diversion device, we examined the effects of stemflow reduction of 25% and 50% on the growth and leaf traits of two common but distinct epiphyte species in a subtropical forest. One species (Asplenium nidus) has a large substrate, and another (Haplopteris zosterifolia) without known water storage structure. The 25% stemflow reduction had limited effects on the growth and leaf traits of the two epiphytes, indicating that 25% stemflow reduction did not reach the water stress threshold for the two epiphytes. It is also possible that the actual reduction in water availability was less than 25% since epiphytes do not use all available stemflow and there could be other sources of water such as fog and throughfall. The 50% stemflow reduction reduced leaf number and leaf area of H. zosterifolia, but not A. nidus, likely because water stored in the large substrate of A. nidus mitigated the impact of stemflow reduction. The thinner leaves, smaller leaf dry matter content, and lower δ13C of A. nidus than H. zosterifolia support the role of water storage of the substrate of A. nidus on mitigating water stress. The 50% stemflow reduction increased leaf thickness, leaf dry matter content of H. zosterifolia, and thickness of abaxial cuticle layer of both epiphytes but had no effect on δ13C. Stemflow reduction had no effects on nutrient concentration and nutrient ratios of both epiphytes suggesting that the epiphytes were able to maintain stoichiometry. Our results indicate that the epiphytes minimized nonstomatal water loss when environmental dryness increased but maintained stomata conductance, which could be important in minimizing the impacts of drought on plant growth and quickly resuming growth once drought ends. Our study highlights that not all epiphytes are similarly vulnerable to drought and precipitation reduction may change the relative abundance of epiphytes with and without water storage structure.

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.

Leaf epicuticular wax as a determinant trait for drought resistance in cereals

The wax coating is an element of the lipid protective layer, the cuticle, which covers the above-ground organs of plants and is the main barrier that prevents non-stomatal water loss. The cuticle helps protect plant surfaces from pathogens and ultraviolet radiation and influences interactions between plants and insects. The accumulation of cuticular wax is one of the mechanisms of adaptation to drought stress. As a result, there are more and more reports on the relationship between cuticular wax and plant resistance to drought. This article deals with the relationship between cuticular waxes and drought resistance in cereals. The paper presents the achievements to date on (i) the relationship between wax biosynthesis and plant response to drought stress using glaucous and nonglaucous near-isogenic lines and on (ii) investigating the role of genes for cuticular wax biosynthesis and transport and the transcription factors that regulate them using mutants with disrupted wax coat formation.

Leaf cuticular waxes of wild-type Welsh onion (Allium fistulosum L.) and a wax-deficient mutant: Compounds with terminal and mid-chain functionalities

Plant cuticles cover aerial organs to limit non-stomatal water loss and protect against insects and pathogens. Cuticles contain complex mixtures of fatty acid-derived waxes, with various chain lengths and diverse functional groups. To further our understanding of the chemical diversity and biosynthesis of these compounds, this study investigated leaf cuticular waxes of Welsh onion (Allium fistulosum L.) wild type and a wax-deficient mutant. Leaf waxes were extracted with chloroform, separated using thin layer chromatography (TLC), and analyzed using gas chromatography-mass spectrometry (GC-MS). The extracts contained typical wax compound classes found in nearly all plant lineages but also two uncommon compound classes. Analyses of characteristic MS fragmentation patterns followed by comparisons with synthetic standards identified the latter as very-long-chain ketones and primary ketols. The ketols were minor compounds, with chain lengths ranging from C28 to C32 and carbonyls mainly on C-18 and C-20 in wild type wax, and a C28 chain with C-16 carbonyl in the mutant. The ketones made up 70% of total wax in the wild type, consisting mainly of C31 isomers with carbonyl group on C-14 or C-16. In contrast, the mutant wax comprised only 4% ketones, with chain lengths C27 and C29 and carbonyls predominantly on C-12 and C-14, respectively. A two-carbon homolog shift between wild type and mutant was also observed in the primary alcohols (a major wax compound class), whilst alkanes exhibited a four-carbon shift. Overall, the compositional data shed light on possible biosynthetic pathways to wax ketones that can be tested in future studies.

Suppression of cuticular wax biosynthesis mediated by rice LOV KELCH REPEAT PROTEIN 2 supports a negative role in drought stress tolerance.

Drought tolerance is important for grain crops, including rice (Oryza sativa); for example, rice cultivated under intermittent irrigation produces less methane gas compared to rice grown in anaerobic paddy field conditions, but these plants require greater drought tolerance. Moreover, the roles of rice circadian-clock genes in drought tolerance remain largely unknown. Here, we show that the mutation of LOV KELCH REPEAT PROTEIN 2 (OsLKP2) enhanced drought tolerance by increasing cuticular wax biosynthesis. Among ZEITLUPE family genes, OsLKP2 expression specifically increased under dehydration stress. OsLKP2 knockdown (oslkp2-1) and knockout (oslkp2-2) mutants exhibited enhanced drought tolerance. Cuticular waxes inhibit non-stomatal water loss. Under drought conditions, total wax loads on the leaf surface increased by approximately 10% in oslkp2-1 and oslkp2-2 compared to the wild type, and the transcript levels of cuticular wax biosynthesis genes were upregulated in the oslkp2 mutants. Yeast two-hybrid, bimolecular fluorescence complementation, and coimmunoprecipitation assays revealed that OsLKP2 interacts with GIGANTEA (OsGI) in the nucleus. The osgi mutants also showed enhanced tolerance to drought stress, with a high density of wax crystals on their leaf surface. These results demonstrate that the OsLKP2-OsGI interaction negatively regulates wax accumulation on leaf surfaces, thereby decreasing rice resilience to drought stress.

Characterization of Glossy Spike Mutants and Identification of Candidate Genes Regulating Cuticular Wax Synthesis in Barley (Hordeum vulgare L.).

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

Mutation of OsMYB60 reduces rice resilience to drought stress by attenuating cuticular wax biosynthesis.

The cuticular wax layer on leaf surfaces limits non-stomatal water loss to the atmosphere and protects against pathogen invasion. Although many genes associated with wax biosynthesis and wax transport in plants have been identified, their regulatory mechanisms remain largely unknown. Here, we show that the MYB transcription factor OsMYB60 positively regulates cuticular wax biosynthesis and this helps rice (Oryza sativa) plants tolerate drought stress. Compared with the wild type (japonica cultivar 'Dongjin'), osmyb60 null mutants (osmyb60-1 and osmyb60-2) exhibited increased drought sensitivity, with more chlorophyll leaching and higher rates of water loss. Quantitative reverse-transcription PCR showed that the loss of function of OsMYB60 led to downregulation of wax biosynthesis genes, leading to reduced amounts of total wax components on leaf surfaces under normal conditions. Yeast one-hybrid, luciferase transient transcriptional activity, and chromatin immunoprecipitation assays revealed that OsMYB60 directly binds to the promoter of OsCER1 (a key gene involved in very-long-chain alkane biosynthesis) and upregulates its expression. Taken together, these results demonstrate that OsMYB60 enhances rice resilience to drought stress by promoting cuticular wax biosynthesis on leaf surfaces.

Cuticular Wax Modification by Epichloë Endophyte in Achnatherum inebrians under Different Soil Moisture Availability.

The cuticular wax serves as the outermost hydrophobic barrier of plants against nonstomatal water loss and various environmental stresses. An objective of this study was to investigate the contribution of the mutualistic fungal endophyte Epichloë gansuensis to leaf cuticular wax of Achnatherum inebrians under different soil moisture availability. Through a pot experiment and gas chromatography−mass spectrometry (GC−MS) analysis, our results indicated that the hydrocarbons were the dominant components of leaf cuticular wax, and the proportion of alcohols, aldehydes, amines, and ethers varied with the presence or absence of E. gansuensis and different soil moisture availability. Amines and ethers are unique in endophyte-free (EF) A. inebrians plants and endophyte-infected (EI) A. inebrians plants, respectively. By transcriptome analysis, we found a total of 13 differentially expressed genes (DEGs) related to cuticular biosynthesis, including FabG, desB, SSI2, fadD, BiP, KCS, KAR, FAR, and ABCB1. A model is proposed which provides insights for understanding cuticular wax biosynthesis in the association of A. inebrians plants with E. gansuensis. These results may help guide the functional analyses of candidate genes important for improving the protective layer of cuticular wax of endophyte-symbiotic plants.

Building a Barrier: The Influence of Different Wax Fractions on the Water Transpiration Barrier of Leaf Cuticles.

Waxes are critical in limiting non-stomatal water loss in higher terrestrial plants by making up the limiting barrier for water diffusion across cuticles. Using a differential extraction protocol, we investigated the influence of various wax fractions on the cuticular transpiration barrier. Triterpenoids (TRPs) and very long-chain aliphatics (VLCAs) were selectively extracted from isolated adaxial leaf cuticles using methanol (MeOH) followed by chloroform (TCM). The water permeabilities of the native and the solvent-treated cuticles were measured gravimetrically. Seven plant species (Camellia sinensis, Ficus elastica, Hedera helix, Ilex aquifolium, Nerium oleander, Vinca minor, and Zamioculcas zamiifolia) with highly varying wax compositions ranging from nearly pure VLCA- to TRP-dominated waxes were selected. After TRP removal with MeOH, water permeability did not or only slightly increase. The subsequent VLCA extraction with TCM led to increases in cuticular water permeabilities by up to two orders of magnitude. These effects were consistent across all species investigated, providing direct evidence that the cuticular transpiration barrier is mainly composed of VLCA. In contrast, TRPs play no or only a minor role in controlling water loss.

Tubular epicuticular wax is an important trait for limiting non-stomatal water loss from leaves in several <i>Dianthus</i> species

Tubular epicuticular wax is an important trait for limiting non-stomatal water loss from leaves in several <i>Dianthus</i> species

QTL analysis reveals the effect of CER1-1 and CER1-3 to reduce fruit water loss by increasing cuticular wax alkanes in citrus fruit

Postharvest water loss causes fruit softening, wilting, and a decline in quality and commodity value. Cuticular wax is a key barrier against non-stomatal water loss and plays a crucial role in fruit quality maintenance. However, there has been limited research on the genetic basis of fruit wax and postharvest water loss. Here, we found that HJ (Citrus reticulata) and ZK (Poncirus trifoliata) fruit had significant differences in postharvest water loss and cuticular wax. HJ fruit had a lower wax content and a faster water loss, and the main aliphatic wax components were alkanes and aldehydes. By contrast, ZK fruit had a higher wax content and a slower water loss, and the main aliphatic wax components were only alkanes. Correlation analysis revealed that fruit water loss seemed to be correlated with cuticular wax alkane in the F1 pseudo-testcross population of HJ and ZK. Furthermore, through high-density genetic map and bulk segregant analysis, alkanes and fruit water loss were co-localized to QTL3, resulting in the identification of CER1-1 and CER1-3 as the candidate genes. Heterologous overexpression in Arabidopsis revealed that they were involved in alkane synthesis and reduction of leaf water loss. In addition, coating with C28 alkane could reduce postharvest fruit water loss of six citrus varieties. Collectively, we speculated that cuticular wax alkanes might play an important role in limiting fruit water loss. The study provides new insights into the development of coating agents and breeding of citrus varieties with better storage performance.

The Tea Plant Leaf Cuticle: From Plant Protection to Tea Quality.

Camellia sinensis (tea tree) is a perennial evergreen woody crop that has been planted in more than 50 countries worldwide; its leaves are harvested to make tea, which is one of the most popular nonalcoholic beverages. The cuticle is the major transpiration barrier to restrict nonstomatal water loss and it affects the drought tolerance of tea plants. The cuticle may also provide molecular cues for the interaction with herbivores and pathogens. The tea-making process almost always includes a postharvest withering treatment to reduce leaf water content, and many studies have demonstrated that withering treatment-induced metabolite transformation is essential to shape the quality of the tea made. Tea leaf cuticle is expected to affect its withering properties and the dynamics of postharvest metabolome remodeling. In addition, it has long been speculated that the cuticle may contribute to the aroma quality of tea. However, concrete experimental evidence is lacking to prove or refute this hypothesis. Even though its relevance to the abiotic and biotic stress tolerance and postharvest processing properties of tea tree, tea cuticle has long been neglected. Recently, there are several studies on the tea cuticle regarding its structure, wax composition, transpiration barrier organization, environmental stresses-induced wax modification, and structure–function relations. This review is devoted to tea cuticle, the recent research progresses were summarized and unresolved questions and future research directions were also discussed.