Non-stomatal Water Loss Research Articles

A co-opted steroid synthesis gene, maintained in sorghum but not maize, is associated with a divergence in leaf wax chemistry

Virtually all land plants are coated in a cuticle, a waxy polyester that prevents nonstomatal water loss and is important for heat and drought tolerance. Here, we describe a likely genetic basis for a divergence in cuticular wax chemistry between Sorghum bicolor, a drought tolerant crop widely cultivated in hot climates, and its close relative Zea mays (maize). Combining chemical analyses, heterologous expression, and comparative genomics, we reveal that: 1) sorghum and maize leaf waxes are similar at the juvenile stage but, after the juvenile-to-adult transition, sorghum leaf waxes are rich in triterpenoids that are absent from maize; 2) biosynthesis of the majority of sorghum leaf triterpenoids is mediated by a gene that maize and sorghum both inherited from a common ancestor but that is only functionally maintained in sorghum; and 3) sorghum leaf triterpenoids accumulate in a spatial pattern that was previously shown to strengthen the cuticle and decrease water loss at high temperatures. These findings uncover the possibility for resurrection of a cuticular triterpenoid-synthesizing gene in maize that could create a more heat-tolerant water barrier on the plant's leaf surfaces. They also provide a fundamental understanding of sorghum leaf waxes that will inform efforts to divert surface carbon to intracellular storage for bioenergy and bioproduct innovations.

Dissecting the Roles of Cuticular Wax in Plant Resistance to Shoot Dehydration and Low-Temperature Stress in Arabidopsis.

Cuticular waxes are a mixture of hydrophobic very-long-chain fatty acids and their derivatives accumulated in the plant cuticle. Most studies define the role of cuticular wax largely based on reducing nonstomatal water loss. The present study investigated the role of cuticular wax in reducing both low-temperature and dehydration stress in plants using Arabidopsis thaliana mutants and transgenic genotypes altered in the formation of cuticular wax. cer3-6, a known Arabidopsis wax-deficient mutant (with distinct reduction in aldehydes, n-alkanes, secondary n-alcohols, and ketones compared to wild type (WT)), was most sensitive to water loss, while dewax, a known wax overproducer (greater alkanes and ketones compared to WT), was more resistant to dehydration compared to WT. Furthermore, cold-acclimated cer3-6 froze at warmer temperatures, while cold-acclimated dewax displayed freezing exotherms at colder temperatures compared to WT. Gas Chromatography-Mass Spectroscopy (GC-MS) analysis identified a characteristic decrease in the accumulation of certain waxes (e.g., alkanes, alcohols) in Arabidopsis cuticles under cold acclimation, which was additionally reduced in cer3-6. Conversely, the dewax mutant showed a greater ability to accumulate waxes under cold acclimation. Fourier Transform Infrared Spectroscopy (FTIR) also supported observations in cuticular wax deposition under cold acclimation. Our data indicate cuticular alkane waxes along with alcohols and fatty acids can facilitate avoidance of both ice formation and leaf water loss under dehydration stress and are promising genetic targets of interest.

Molecular, chemical, and physiological analyses of sorghum leaf wax under post-flowering drought stress

Wax accumulation on the sorghum surface plays an important role in drought tolerance by preventing non-stomatal water loss. Thereby, the effect of post-flowering drought stress (PFDS) on the epicuticular wax (EW) amount, relative water content (RWC), chlorophyll, and grain yield in sorghum drought contrasting genotypes were investigated. The experiment was conducted as a split-plot based on randomized complete block design (RCBD) with two water treatments (normal watering and water holding after 50% flowering stage), and three genotypes (Kimia and KGS23 as drought-tolerant and Sepideh as drought-susceptible). Scanning electron microscopy and GC-MS analyses were used to determine the wax crystals density and its compositions, respectively. In addition, based on literature reviews and publicly available datasets, six wax biosynthesis drought stress-responsive genes were chosen for expression analysis. The results showed that the amounts of EW and wax crystals density were increased in Kimia and Sepideh genotypes and no changed in KGS23 genotype under drought stress. Chemical compositions of wax were classified into six major groups including alkanes, fatty acids, aldehydes, esters, alcohols, and cyclic compounds. Alkanes increment in drought-tolerant genotypes led to make an effective barrier against the drought stress to control water losses. In addition, the drought-tolerant genotypes had higher levels of RWC compared to the drought-susceptible ones, resulted in higher yield produced under drought condition. According to the results, SbWINL1, FATB, and CER1 genes play important roles in drought-induced wax biosynthesis. The results of the present study revealed a comprehensive view of the wax and its compositions and some involved genes in sorghum under drought stress.

Drought-Responsive ZmFDL1/MYB94 Regulates Cuticle Biosynthesis and Cuticle-Dependent Leaf Permeability.

In all land plants, the outer surface of aerial parts is covered by the cuticle, a complex lipid layer that constitutes a barrier against damage caused by environmental factors and provides protection against nonstomatal water loss. We show in this study that both cuticle deposition and cuticle-dependent leaf permeability during the juvenile phase of plant development are controlled by the maize (Zea mays) transcription factor ZmFUSED LEAVES 1 (FDL1)/MYB94. Biochemical analysis showed altered cutin and wax biosynthesis and deposition in fdl1-1 mutant seedlings at the coleoptile stage. Among cutin compounds, ω-hydroxy fatty acids and polyhydroxy-fatty acids were specifically affected, while the reduction of epicuticular waxes was mainly observed in primary long chain alcohols and, to a minor extent, in long-chain wax esters. Transcriptome analysis allowed the identification of candidate genes involved in lipid metabolism and the assembly of a proposed pathway for cuticle biosynthesis in maize. Lack of ZmFDL1/MYB94 affects the expression of genes located in different modules of the pathway, and we highlighted the correspondence between gene transcriptional variations and biochemical defects. We observed a decrease in cuticle-dependent leaf permeability in maize seedlings exposed to drought as well as abscisic acid treatment, which implies coordinated changes in the transcript levels of ZmFDL1/MYB94 and associated genes. Overall, our results suggest that the response to water stress implies the activation of wax biosynthesis and the involvement of both ZmFDL1/MYB94 and abscisic acid regulatory pathways.

Wax biosynthesis in response to danger: its regulation upon abiotic and biotic stress.

The plant cuticle is the first physical barrier between land plants and their terrestrial environment. It consists of the polyester scaffold cutin embedded and sealed with organic, solvent-extractable cuticular waxes. Cuticular wax ultrastructure and chemical composition differ with plant species, developmental stage and physiological state. Despite this complexity, cuticular wax consistently serves a critical role in restricting nonstomatal water loss. It also protects the plant against other environmental stresses, including desiccation, UV radiation, microorganisms and insects. Within the broader context of plant responses to abiotic and biotic stresses, our knowledge of the explicit roles of wax crystalline structures and chemical compounds is lacking. In this review, we summarize our current knowledge of wax biosynthesis and regulation in relation to abiotic and biotic stresses and stress responses.

Phosphatase AtDBP1 negatively regulates drought and salt tolerance through altering leaf surface permeability in Arabidopsis.

In our previous study, AtDBP1 encoding a DBP factor was identified as a putative abiotic stress candidate gene. DBP factors are important regulators that participate in both transcriptional regulation and post-translational regulation, but their roles in abiotic stress are still not well-understood. So we conducted a detailed study on the function of AtDBP1 in abiotic stress. It is found that expression of AtDBP1 could be induced by drought and salt, and the induction by salt was inhibited in ABA-deficient mutant aba2-3, indicating the expression of AtDBP1 was ABA-inducible. Overexpression of AtDBP1 resulted in a rapid stomatal closure, and elevated expression of drought/salt-responsive genes, which should help Arabidopsis to enhance the drought and salt tolerance. Unexpectedly, overexpression of AtDBP1 decreased the drought and salt tolerance of Arabidopsis. Further analysis suggested that AtDBP1 is involved in cuticle wax and cuticle membrane regulation. Overexpression of AtDBP1 showed increased cuticular conductance due to a decreased cuticle wax accumulation and cuticle membrane thickness. The cuticular wax provides an essential barrier for decreasing nonstomatal water loss during drought stress, so overexpression of AtDBP1 showed decreased drought tolerance possibly ascribed to the change of cuticle membrane structure. Our previous study elucidated that AtDBP1 was also involved in flowering time regulation. Taken together, the results above indicated that AtDBP1 was involved in both plant development and stress regulation. The mechanism of AtDBP1 in this study indicates that genes involved in both plant development and stress regulation might be not suitable for production application in breeding. Collectively, our results provide some new ideas on purposefully increasing the abiotic stress without influence on plant growth and development.

Drought stress modify cuticle of tender tea leaf and mature leaf for transpiration barrier enhancement through common and distinct modes

Cuticle is the major transpiration barrier that restricts non-stomatal water loss and is closely associated with plant drought tolerance. Although multiple efforts have been made, it remains controversial what factors shape up the cuticular transpiration barrier. Previously, we found that the cuticle from the tender tea leaf was mainly constituted by very-long-chain-fatty-acids and their derivatives while alicyclic compounds dominate the mature tea leaf cuticle. The presence of two contrasting cuticle within same branch offered a unique system to investigate this question. In this study, tea seedlings were subjected to water deprivation treatment, cuticle structures and wax compositions from the tender leaf and the mature leaf were extensively measured and compared. We found that cuticle wax coverage, thickness, and osmiophilicity were commonly increased from both leaves. New waxes species were specifically induced by drought; the composition of existing waxes was remodeled; the chain length distributions of alkanes, esters, glycols, and terpenoids were altered in complex manners. Drought treatment significantly reduced leaf water loss rates. Wax biosynthesis-related gene expression analysis revealed dynamic expression patterns dependent on leaf maturity and the severity of drought. These data suggested that drought stress-induced structural and compositional cuticular modifications improve cuticle water barrier property. In addition, we demonstrated that cuticle from the tender leaf and the mature leaf were modified through both common and distinct modes.

Alkanes (C29 and C31)-Mediated Intracuticular Wax Accumulation Contributes to Melatonin- and ABA-Induced Drought Tolerance in Watermelon

As the outermost hydrophobic layer, cuticular waxes serve as an essential waterproof barrier to protect plants from desiccation, but the mechanism of wax accumulation still remains unclear. We analyzed the response of cuticular wax composition and deposition to drought in three different watermelon germplasms, namely, M20, M08, and J5F, which showed similar stomatal response, but high, moderate, and low tolerance to drought, respectively. Among the identified 28 compounds of cuticular waxes on leaves, more alkanes with chain lengths of C29 and C31 were induced in M20, accompanied by an increased transcript levels of CER1 (very-long-chain aldehyde decarbonylase 1), when compared to that in M08 and J5F. M20 showed higher total wax amount but fewer platelet-like wax crystals on the upper epidermis of leaves under drought, suggesting the prevalence of more intracuticular waxes embedded into the cutin matrix as fillers. These distinct responses of cuticular waxes in M20 conferred low water loss and high tolerance to drought. Melatonin and abscisic acid (ABA), which can be induced by drought, promoted the biosynthesis of alkanes (C29 and C31) but inhibited the accumulation of wax crystals under drought. Moreover, melatonin inhibited the elevation of ABA levels under mild drought but promoted the ABA accumulation under severe drought, indicating that melatonin and ABA function synergistically to regulate wax compositions to limit non-stomatal water loss under severe but not mild drought in watermelon. These findings can be exploited to improve crop tolerance to drought in arid regions.

Transcriptomic and Gas Chromatography-Mass Spectrometry Metabolomic Profiling Analysis of the Epidermis Provides Insights into Cuticular Wax Regulation in Developing 'Yuluxiang' Pear Fruit.

The layer of cuticular wax covering fruits plays important roles in protecting against disease, preventing non-stomatal water loss, and extending shelf life. However, the molecular basis of cuticular wax biosynthesis in pear (Pyrus) fruits remains elusive. Our study thoroughly investigates cuticular wax biosynthesis during pear fruit development from morphologic, transcriptomic, and gas chromatography-mass spectrometry metabolomic perspectives. Our results showed that cuticular wax concentrations increased during the early stage [20-80 days after full bloom (DAFB)] from 0.64 mg/cm2 (50 DAFB) to 1.75 mg/cm2 (80 DAFB) and then slightly decreased to 1.22 mg/cm2 during the fruit ripening period (80-140 DAFB). Scanning electron microscopy imaging indicated that wax plate crystals increased and wax structures varied during the pear fruit development. The combined transcriptomic and metabolomic profiling analysis revealed 27 genes, including 12 genes encoding transcription factors and a new structural gene (Pbr028523) encoding β-amyrin synthase, participating in the biosynthesis, transport, and regulation of cuticular wax according to their expression patterns in pear fruit. The quantitative real-time polymerase chain reaction experiments of 18 differentially expressed genes were performed and confirmed the accuracy of the RNA-Seq-derived transcript expression. A model of VLCFAs and cuticular wax synthesis and transport in pear fruit is proposed, providing a mechanistic framework for understanding cuticular wax biosynthesis in pear fruit. These results and data sets provide a foundation for the molecular events related to cuticular wax in 'Yuluxiang' pear fruit and may also help guide the functional analyses of candidate genes important for improving the cuticular wax of pear fruit in the future.

Developmental and Environmental Regulation of Cuticular Wax Biosynthesis in Fleshy Fruits.

The aerial parts of land plants are covered by a hydrophobic layer called cuticle that limits non-stomatal water loss and provides protection against external biotic and abiotic stresses. The cuticle is composed of polymer cutin and wax comprising a mixture of very-long-chain fatty acids and their derivatives, while also bioactive secondary metabolites such as triterpenoids are present. Fleshy fruits are also covered by the cuticle, which has an important protective role during the fruit development and ripening. Research related to the biosynthesis and composition of cuticles on vegetative plant parts has largely promoted the research on cuticular waxes in fruits. The chemical composition of the cuticular wax varies greatly between fruit species and is modified by developmental and environmental cues affecting the protective properties of the wax. This review focuses on the current knowledge of the cuticular wax biosynthesis during fleshy fruits development, and on the effect of environmental factors in regulation of the biosynthesis. Bioactive properties of fruit cuticular waxes are also briefly discussed, as well as the potential for recycling of industrial fruit residues as a valuable raw material for natural wax to be used in food, cosmetics and medicine.

Variation in leaf wax n-alkane characteristics with climate in the broad-leaved paperbark (Melaleuca quinquenervia)

In higher plants, leaf waxes provide a barrier to non-stomatal water loss, and their composition varies both between and within species. Characteristics of n-alkanes, a suite of ubiquitous compounds in these waxes, are thought to be influenced by the availability of water and the temperature in a plant’s growing environment. Longer n-alkane distributions with less variability in chain length are hypothesised to confer greater resistance to non-stomatal water loss and thus are expected in higher abundance in desiccating environments. Relationships between the distribution of n-alkanes and both precipitation and temperature have previously been observed. Despite this, it is unclear whether n-alkane chain length distributions vary plastically in response to climate, or whether they are fixed within populations in different climate settings. To better understand this, we examine the relationship between n-alkane characteristics of Melaleuca quinquenervia and both spatial and temporal climate variation. Across eastern Australia, we find that n-alkane homolog concentrations and distributions in leaves of M. quinquenervia do not vary with climate where samples are proximate, even when climate shows significant variability. However, the concentration and distribution of n-alkane homologs do differ considerably between geographically separated populations in very different climate regimes. These results suggest n-alkane characteristics are not a plastic response to climate variability, and instead are likely fixed and could be driven by genetic differences between populations. This has important implications for the use of n-alkane characteristics as palaeoenvironmental proxies.

Overexpression of ß-Ketoacyl Co-A Synthase1 Gene Improves Tolerance of Drought Susceptible Groundnut (Arachis hypogaea L.) Cultivar K-6 by Increased Leaf Epicuticular Wax Accumulation.

Drought is one of the major environmental constraints affecting the crop productivity worldwide. One of the agricultural challenges today is to develop plants with minimized water utilization and reduced water loss in adverse environmental conditions. Epicuticular waxes play a major role in minimizing water loss. Epicuticular wax covers aerial plant parts and also prevents non-stomatal water loss by forming the outermost barrier from the surfaces. Epicuticular wax content (EWC) variation was found to be affiliated with drought tolerance of groundnut cultivars. In the current study, a fatty acid elongase gene, KCS1, which catalyzes a rate limiting step in the epicuticular wax biosynthesis was isolated from drought tolerant cultivar K-9 and overexpressed in drought sensitive groundnut cultivar (K-6) under the control of CaMV35S constitutive promoter. Transgenic groundnut plants overexpressing AhKCS1 exhibited normal growth and displaying greenish dark shiny appearance. Environmental scanning electron microscopy (ESEM) revealed the excess of epicuticular wax crystal depositions on the transgenic plant leaves compared to non-transgenic wild type plants. The findings were further supported by gas chromotography–mass spectroscopic analysis (GC-MS) that revealed enhanced levels of fatty acids, secondary alcohols, primary alcohols, aldehydes, alkanes, and ketones in transgenics compared to wild types. The AhKCS1 overexpressing transgenic groundnut plants exhibited increase in the cuticular wax content, reduction of water loss, lower membrane damage, decreased MDA content, and high proline content compared to that of non-transgenic groundnut plants. Our findings suggest that the AhKCS1 gene plays a major role in combating drought stress by preventing non-stomatal water loss in drought sensitive groundnut cultivar (K-6).

OsABCG9 Is an Important ABC Transporter of Cuticular Wax Deposition in Rice.

The importance of the cuticular layer in regulating a plant’s water status and providing protection from environmental challenges has been recognized for a long time. The cuticular layer in plants restricts non-stomatal water loss and protects plants against damage from pathogen infection and UV radiation. Much genetic and biochemical research has been done about cutin and wax transportation in Arabidopsis thaliana, but little is known about it in rice. Here, we report that a rice ATP-binding cassette (ABC) transporter, OsABCG9, is essential for normal development during vegetative growth and could play a critical role in the transportation of epicuticular wax in rice. Rice phenotypes with mutated OsABCG9 exhibited growth retardation and sensitivity to low humidity. The total amount of cuticular wax on the leaves of the osabcg9-1 mutant diminished by 53% compared with the wild type, and wax crystals disappeared completely in osabcg9-2 mutant leaves. However, OsABCG9 does not seem to be involved in cutin transportation, even though its ortholog in Arabidopsis, AtABCG11, transports both wax and cutin. Furthermore, the osabcg9-1 mutant had increased leaf chlorophyll leaching and more severe drought susceptibility. This study provides new insights about differences between rice and A. thaliana in wax and cutin transportation associated with the ABCG family during evolution.

Exogenous Melatonin Improves Tolerance to Water Deficit by Promoting Cuticle Formation in Tomato Plants.

The plant cuticle, composed of cutin and waxes, is a hydrophobic layer coating the aerial organs of terrestrial plants and playing a critical role in limiting water loss. While melatonin has been recently demonstrated to be involved in responses to drought stress in plants, its relationship with cuticle formation is not known. In the present work, we report the effects of melatonin on the formation of cuticle in tomato leaves subjected to water deficit. Preliminary analysis by light microscope showed that tomato leaves pretreated with exogenous melatonin might have thicker cutin than tomato leaves without melatonin pretreatment under water deficit condition. Chemical characterization showed that exogenous application of melatonin increased the level of cuticular waxes in tomato leaves under water deficit. Consistent with the change in cuticular waxes was the increased abundance of wax-associated gene transcripts. Further, assessment of water loss and chlorophyll leaching in tomato leaves revealed the association of cuticle deposition with reduced leaf permeability, which is important in restricting water loss in water deficit-stressed tomato plants. These results suggest a role for melatonin in regulating leaf cuticle formation and non-stomatal water loss in leaves.

Three endoplasmic reticulum-associated fatty acyl-coenzyme a reductases were involved in the production of primary alcohols in hexaploid wheat (Triticum aestivum L.)

BackgroundThe cuticle covers the surface of the polysaccharide cell wall of leaf epidermal cells and forms an essential diffusion barrier between the plant and the environment. The cuticle is composed of cutin and wax. Cuticular wax plays an important role in the survival of plants by serving as the interface between plants and their biotic and abiotic environments, especially restricting nonstomatal water loss. Leaf cuticular waxes of hexaploid wheat at the seedling stage mainly consist of primary alcohols, aldehydes, fatty acids, alkane and esters. Primary alcohols account for more than 80% of the total wax load. Therefore, we cloned several genes encoding fatty acyl-coenzyme A reductases from wheat and analyzed their function in yeast and plants. We propose the potential use of these genes in wheat genetic breeding.ResultsWe reported the cloning and characterization of three TaFARs, namely TaFAR6, TaFAR7 and TaFAR8, encoding fatty acyl-coenzyme A reductases (FAR) in wheat leaf cuticle. Expression analysis revealed that TaFAR6, TaFAR7 and TaFAR8 were expressed at the higher levels in the seedling leaf blades, and were expressed moderately or weakly in stamen, glumes, peduncle, flag leaf blade, sheath, spike, and pistil. The heterologous expression of three TaFARs in yeast (Saccharomyces cerevisiae) led to the production of C24:0 and C26:0 primary alcohols. Transgenic expression of the three TaFARs in tomato (Solanum lycopersicum) and rice (Oryza sativa) led to increased accumulation of C24:0–C30:0 primary alcohols. Transient expression of GFP protein-tagged TaFARs revealed that the three TaFAR proteins were localized to the endoplasmic reticulum (ER), the site of wax biosynthesis. The three TaFAR genes were transcriptionally induced by drought, cold, heat, powdery mildew (Blumeria graminis) infection, abscisic acid (ABA) and methyl jasmonate (MeJa) treatments.ConclusionsThese results indicated that wheat TaFAR6, TaFAR7 and TaFAR8 are involved in biosynthesis of very-long-chain primary alcohols in hexaploid wheat and in response to multiple environmental stresses.

Characterization of leaf cuticular wax classes and constituents in a spring Camelina sativa diversity panel

Among oilseed species, Camelina has received considerable attention as an oilseed crop that can be manipulated easily to meet important non-food bioenergy requirements, where it is relatively high in oil content and polyunsaturated fatty acids, and has a very short growing season with fairly good adaption to marginal lands and low input agricultural systems. To expand Camelina cultivation zones into more arid regions, it is important to develop new drought resistant cultivars that can grow under water-limited conditions. Increasing accumulated leaf cuticular wax in Camelina could be one of the strategies to reduce nonstomatal water loss and thus increase crop tolerance to drought. To extend the understanding of phenotypic variations in cuticular wax content and composition in Camelina sativa, leaf wax constituents from a spring Camelina diversity panel containing 163 accessions were extracted and analyzed. The diversity panel exhibited a wide range in total leaf wax contents, wax classes and constituents. Among primary alcohols, the dominant constituents were the C24, C26 and C28 homologues, while the C31 homologue was the most abundant alkane among all Camelina accessions. High heritability values of the primary alcohol class and its dominant constituent C24, C26 and C28 homologues, as well as the alkane class and abundant C29, C31, and C33 constituents, suggested the feasibility for selection of these traits during early generations of Camelina breeding programs. Positive correlations among leaf wax content, wax classes and their constituents suggest that modifying specific wax constituents could increase the wax loads, which in turn could enhance cuticle composition and properties. Quantification of leaf wax traits in the Camelina diversity panel will underpin future analysis of the Camelina wax biosynthetic pathways, help dissect its genetic regulatory elements, identify candidate genes controlling these traits, and enable the development of molecular markers for molecular breeding programs aimed at increasing drought tolerance of Camelina.

Cloning of the Brcer1 gene involved in cuticular wax production in a glossy mutant of non-heading Chinese cabbage (Brassica rapa L. var. communis)

Cuticular wax is a complex mixture of very-long-chain fatty acid derivatives. The wax on the surface of plants serves as a protective barrier to reduce non-stomatal water loss and environmental damage. However, the loss of wax may lead to a glossy phenotype, which is an favorable trait in leafy vegetables. The mechanism of glossy mutants in non-heading Chinese cabbage (Brassica rapa L. var. communis) has not been studied yet. In this study, scanning electron microscopy (SEM) showed that the cuticular wax on the leaves and stem of a glossy mutant was dramatically reduced compared with that of the wild-type plant. Transmission electron microscopy (TEM) revealed that the cuticle ultrastructure of glossy mutant leaf and stem were altered when compared with the wild type. A cuticle wax analysis showed the total wax content of leaves, as well as alkanes, ketones and alcohols, was decreased. A genetic analysis indicated that the glossy phenotype was controlled by a single gene. Based on a homology analysis, the Brcer1 gene was identified as the candidate gene controlling the glossy phenotype. In the glossy mutant, a 39-bp deletion leads to an mRNA disruption and reduces the expression of the BrCER1 gene. Sequence analysis showed that a loss of function mutation in the Brcer1 gene was different from that of Cgl1, which was previously shown to be responsible for the glossy phenotype in B. oleracea, showing typical parallel selection. These findings provide a better understanding of the cuticular wax biosynthesis pathway and offer important information for molecular-assisted breeding of non-heading Chinese cabbage (B. rapa L. var. communis).

The Polyketide Components of Waxes and the Cer-cqu Gene Cluster Encoding a Novel Polyketide Synthase, the β-Diketone Synthase, DKS.

The primary function of the outermost, lipophilic layer of plant aerial surfaces, called the cuticle, is preventing non-stomatal water loss. Its exterior surface is often decorated with wax crystals, imparting a blue–grey color. Identification of the barley Cer-c, -q and -u genes forming the 101 kb Cer-cqu gene cluster encoding a novel polyketide synthase—the β-diketone synthase (DKS), a lipase/carboxyl transferase, and a P450 hydroxylase, respectively, establishes a new, major pathway for the synthesis of plant waxes. The major product is a β-diketone (14,16-hentriacontane) aliphatic that forms long, thin crystalline tubes. A pathway branch leads to the formation of esterified alkan-2-ols.

Control of Plant Water Use by ABA Induction of Senescence and Dormancy: An Overlooked Lesson from Evolution

Drought stress is a condition that in specific climate contexts results in insufficient water availability and often limits plant productivity through perturbing development and reducing plant growth and survival. Plants use senescence of old leaves and dormancy of buds and seeds to survive extreme environmental conditions. The plant hormone ABA accumulates after drought stress, and increases plant survival by inducing quick responses such as stomatal closure, and long-term responses such as extended growth inhibition, osmotic regulation, accumulation of cuticular wax, senescence, abscission and dormancy. Here we focus on how the long-term ABA responses contribute to plant survival during severe drought stress. Leaf senescence and abscission of older leaves reduce total plant transpirational water loss and increase the transfer of nutrients to meristems and to some storage tissues. Osmotic regulation favors water consumption in sink tissues, and accumulation of cuticular wax helps to seal the plant surface and limits non-stomatal water loss.

Five Fatty Acyl-Coenzyme A Reductases Are Involved in the Biosynthesis of Primary Alcohols in Aegilops tauschii Leaves.

The diploid Aegilops tauschii is the D-genome donor to hexaploid wheat (Triticum aestivum) and represents a potential source for genetic study in common wheat. The ubiquitous wax covering the aerial parts of plants plays an important role in protecting plants against non-stomatal water loss. Cuticular waxes are complex mixtures of very-long-chain fatty acids, alkanes, primary and/or secondary alcohols, aldehydes, ketones, esters, triterpenes, sterols, and flavonoids. In the present work, primary alcohols were identified as the major components of leaf cuticular wax in Ae. tauschii, with C26:0-OH being the dominant primary alcohol. Analysis by scanning electron microscope revealed that dense platelet-shaped wax crystals were deposited on leaf surfaces of Ae. tauschii. Ten putative wax biosynthetic genes encoding fatty acyl-coenzyme A reductase (FAR) were identified in the genome of Ae. tauschii. Five of these genes, Ae.tFAR1, Ae.tFAR2, Ae.tFAR3, Ae.tFAR4, and Ae.tFAR6, were found expressed in the leaf blades. Heterologous expression of the five Ae.tFARs in yeast (Saccharomyces cerevisiae) showed that Ae.tFAR1, Ae.tFAR2, Ae.tFAR3, Ae.tFAR4, and Ae.tFAR6 were predominantly responsible for the accumulation of C16:0, C18:0, C26:0, C24:0, and C28:0 primary alcohols, respectively. In addition, nine Ae.tFAR paralogous genes were located on D chromosome of wheat and the wheat nullisomic–tetrasomic lines with the loss of Ae.tFAR3 and Ae.tFAR4 paralogous genes had significantly reduced levels of primary alcohols in the leaf blades. Collectively, these data suggest that Ae.tFAR1, Ae.tFAR2, Ae.tFAR3, Ae.tFAR4, and Ae.tFAR6 encode alcohol-forming FARs involved in the biosynthesis of primary alcohols in the leaf blades of Ae. tauschii. The information obtained in Ae. tauschii enables us to better understand wax biosynthesis in common wheat.