植物角质层蜡质的化学组成研究综述

角质层是植物与外界的第一接触面,而角质层蜡质则是由位于角质层外的外层蜡质和深嵌在角质层中的内层蜡质两部分构成。植物角质层蜡质成分极其复杂,具有重要的生理功能。综述了有关植物角质层蜡质的化学组成信息,探讨了目前植物角质层蜡质化学成分研究中存在的一些问题,展望了角质层蜡质成分的研究前景。     

The positional sterile (ps) mutation affects cuticular transpiration and wax biosynthesis of tomato fruits

Cuticular waxes are known to play a pivotal role in limiting transpirational water loss across primary plant surfaces. The astomatous tomato fruit is an ideal model system that permits the functional characterization of intact cuticular membranes and therefore allows direct correlation of their permeance for water with their qualitative and quantitative composition. The recessive positional sterile (ps) mutation, which occurred spontaneously in tomato (Solanum lycopersicum L.), is characterized by floral organ fusion and positional sterility. Because of a striking phenotypical similarity with the lecer6 wax mutant of tomato, which is defective in very-long-chain fatty acid elongation, ps mutant fruits were analyzed for their cuticular wax and cutin composition. We also examined their cuticular permeance for water following the developmental course of fruit ripening. Wild type and ps mutant fruits showed considerable differences in their cuticular permeance for water, while exhibiting similar quantitative wax accumulation. The ps mutant fruits showed a five- to eightfold increase in water loss per unit time and surface area when compared to the corresponding wild type fruits. The cuticular waxes of ps mutant fruits were characterized by an almost complete absence of n-alkanes and aldehydes, with a concomitant increase in triterpenoids and sterol derivatives. We also noted the occurrence of alkyl esters not present in the wild type. Quantitative and qualitative cutin monomer composition remained largely unaffected. The significant differences in the cuticular wax composition of ps mutant fruits induced a distinct increase of cuticular water permeance. The fruit wax compositional phenotype indicates the ps mutation is responsible for effectively blocking the decarbonylation pathway of wax biosynthesis in epidermal cells of tomato fruits.     

ABC-type transporters and cuticle assembly: Linking function to polarity in epidermis cells

The aerial organs of plants are covered with a cuticle, a continuous layer overlaying the outermost cell walls of the epidermis. The cuticle is composed of two major classes of the lipid biopolymers: cutin and waxes, collectively termed cuticular lipids. Biosynthesis and transport of cuticular lipids occur predominantly in the epidermis cells. In the transport pathway, cuticular lipids are exported from their site of biosynthesis in the ER/plastid to the extracellular space through the plasma membrane and cell wall. Growing evidence suggests that ATP-binding cassette (ABC) transporters are implicated in transport of cuticular lipids across the plasma membrane of epidermal cells. The Arabidopsis ABC-type transporter protein CER5 (WBC12) was reported to act as a wax monomers transporter. In recent works, our group and others showed that a CER5-related protein, DESPERADO (DSO/WBC11), is required for cutin and wax monomers transport through the plasma membrane of Arabidopsis epidermis cells. Unlike the cer5 mutant, DSO loss-of-function had a profound effect on plant growth and development, particularly dwarfism, postgenital organ fusions, and altered epidermal cell differentiation. The partially overlapping function of CER5 and DSO and the fact that these proteins are half-size ABC transporters suggest that they might form a hetero-dimeric complex while transporting wax components. An intriguing observation was the polar localization of DSO in the distal part of epidermis cells. This polar expression might be explained by DSO localization within lipid rafts, specific plasma membrane microdomains which are associated with polar protein expression. In this review we suggest possible mechanisms for cuticular lipids transport and a link between DSO function and polar expression. Furthermore, we also discuss the subsequent transport of cuticular constituents through the hydrophobic cell wall and the possible involvement of lipid transfer proteins in this process.Key words: ABC transporter, cuticular lipids, polar expression, plasma membrane, epidermis     

Cuticular waxes in alpine meadow plants: climate effect inferred from latitude gradient in Qinghai‐Tibetan Plateau

Alpine meadow ecosystems are susceptible to climate changes. Still, climate impact on cuticular wax in alpine meadow plants is poorly understood. Assessing the variations of cuticular wax in alpine meadow plants across different latitudes might be useful for predicting how they may respond to climate change. We studied nine alpine meadows in a climate gradient in the east side of Qinghai‐Tibetan Plateau, with mean annual temperature ranging from −7.7 to 3.2°C. In total, 42 plant species were analyzed for cuticular wax, averaged 16 plant species in each meadow. Only four plant species could be observed in all sampling meadows, including Kobresia humilis,Potentilla nivea,Anaphalis lacteal, and Leontopodium nanum. The amounts of wax compositions and total cuticular wax in the four plant species varied among sampling meadows, but no significant correlation could be observed between them and temperature, precipitation, and aridity index based on plant species level. To analyze the variations of cuticular wax on community level, we averaged the amounts of n‐alkanes, aliphatic acids, primary alcohols, and total cuticular wax across all investigated plant species in each sampling site. The mean annual temperature, mean temperature in July, and aridity index were significantly correlated with the averaged amounts of wax compositions and total cuticular wax. The average chain length of n‐alkanes in both plant and soil linearly increased with increased temperature, whereas reduced with increased aridity index. No significant correlation could be observed between mean annual precipitation and mean precipitation from June to August and the cuticular wax amounts and average chain length. Our results suggest that the survival of some alpine plants in specific environments might be depended on their abilities in adjusting wax deposition on plant leaves, and the alpine meadow plants as a whole respond to climate change, benefiting the stability of alpine meadow ecosystem.     

Intracuticular lipids of spinach leaves

The cuticular wax and cutin components of the cuticular membranes isolated from the leaves of two spinach cultivars have been determined. The membranes contain about 0·007 mg/cm² of cuticular wax which comprises monobasic acids (C₁₆–C₃₈) with hexadecanoic as the major component. The amounts of cutin are comparable with those of cuticular wax and the monomeric constituents are predominantly C₁₈ epoxy compounds. The most abundant monomer is 9,10-epoxy-18-hydroxyoctadecanoic acid (up to 63%) together with substantial amounts of 9,10,18-trihydroxyoctadecanoic acid (up to 22%). Also present are 9,10-epoxyoctadecane-1,18-dioic acid (6–7%) dihydroxyhexadecanoic acid (3–4%) and ω-hydroxymonobasic and fatty acid fractions. The tentative identification of two minor components, 18-hydroxyoxooctadecanoic and 9,10-epoxy-12,18-dihydroxyoctadecanoic acids, is also made. Although spinach membranes have a delicate structure their cutin composition is essentially similar to that of much more substantial membranes.     

Wax glands ultrastructure and chemical composition of wax of giant mealybug Drosicha stebbingii (Green) (Hemiptera: Monophlebidae)

Droschia stebbingii (Green) belonging to the family Monophlebidae (Hemiptera: Coccoidea) is a serious, phytophagous and dimorphic pest. D. stebbingii and D. mangiferae (Green) with sixty-two common hosts and minor morphological variations pose problems towards their taxonomic identification. Light and scanning electron microscopy were used to outline the ultrastructural details of cuticular wax producing structures of D. stebbingii. Three types of wax-pores along with tubular duct were observed on the head, thorax and abdomen. Quinquelocular and multilocular disc-pores exhibited variations in the central loculus with single and double opening. Short curly wax strands were secreted from the outer loculi whereas central loculus secreted long tubular strands. Additionally, bilocular pores secreting curly wax were observed having two large loculi separated by a septum. GC–MS analysis of cuticular wax indicated the presence of hydrocarbons and wax esters forming majority of the covering along with long chain alcohols, ketones and acids. Variations in the percentage of hydrocarbon and wax esters content were observed for two populations of D. stebbingii collected from different hosts. Hydrocarbons were abundant in populations collected on Pongamia pinnata (L.) Pierre whereas long chain esters dominated the wax profile for the population collected from Tectona grandis L.f. Compositional variations in the cuticular wax of D. stebbingii on different host along with knowledge on the ultrastructural wax pores and waxy secretion would provide an insight to the existing knowledge of this scale insect.     

Observation of an anisotropic texture inside the wax layer of insect cuticle

The outermost part of insect cuticles is very often covered with wax, which prevents desiccation and serves for chemical communication in many species. Earlier studies on cuticular waxes have mainly focused on their chemical composition revealing complex mixtures of lipids. In the absence of information on their physical organization, cuticular waxes have been considered isotropic. Here we report the presence of parallel stripes in the wax layer of the carapace of the scarab beetle, Chrysina gloriosa, with a textural periodicity of ca. 28 nm, as revealed by electron microscopy of transverse sections. Observations at oblique incidence argue for a layered organization of the wax, which might be related to a layer-by-layer deposition of excreted wax. Our findings may lay the foundation for further studies on the internal structure of cuticular waxes for other insects.     

空气湿度与土壤水分胁迫对紫花苜蓿叶表皮蜡质特性的影响

选用2个抗旱性不同的紫花苜蓿品种,敖汉(强抗旱)和三得利(弱抗旱),设置空气湿度(45%-55%和75%-85%)和土壤水分胁迫(75%和35%田间持水量)处理,分析紫花苜蓿叶表皮蜡质含量、组分及晶体结构、气体交换参数、水势及脯氨酸含量的变化规律。结果表明,单独土壤水分胁迫时,紫花苜蓿叶表皮蜡质晶体结构及蜡质总量无显著变化;敖汉蜡质组分中烷类、酯类含量增加,醇类含量下降;三得利醇类含量下降,烷类、酯类含量变化不显著。低空气湿度胁迫时,两品种蜡质总量无显著变化,烷类和酯类含量显著增加,醇类含量显著下降,叶表皮片状蜡质晶体结构熔融呈弥漫性,扩大了对叶表面积的覆盖,其蒸腾速率显著低于正常湿度。复合胁迫处理时,叶表皮片状蜡质晶体结构继续呈弥漫性,烷类、酯类、未知蜡质组分含量均高于单独胁迫处理,醇类含量最低,而蜡质总量除三得利显著高于对照外,其余均无显著差异。紫花苜蓿叶表皮蜡质各组分含量(除醇类)及蜡质总量与光合速率呈显著负相关,与蒸腾速率无显著相关关系。蜡质总量与叶水势呈显著正相关。总体上,敖汉蜡质总量显著高于三得利,蜡质组分中烷类物质的增加有助于提高植株的抗旱性。在复合胁迫下,强抗旱品种主要通过气孔因素控制水分散失,而弱抗旱品种通过气孔和非气孔因素共同控制植物水分散失。     

Variations of cuticular wax in mulberry trees and their effects on gas exchange and post-harvest water loss

Though mulberry (Morus alba) tree shows great adaptations to various climate conditions, their leaf water status and photosynthesis are sensitive to climate changes. In the current study, seven widely planted mulberry cultivars in Chongqing, Southwest China, were selected to analyze leaf cuticular wax characteristics, gas exchange index, post-harvest leaf water status and their relationships, aiming to provide new theory in screening high resistant mulberry cultivars. Mulberry trees formed rounded cap-type idioblasts on the adaxial leaf surface. Film-like waxes and granule-type wax crystals covered leaf surfaces, varying in crystal density among cultivars. The stomatal aperture on the abaxial surface of cultivars with high wax amount was smaller than that of cultivars with low wax amount. The amount of total wax was negatively correlated with the net photosynthetic rate (P _N), transpiration rate (E) and stomatal conductance (g _s) and positively correlated with the moisture retention capacity. It suggested that both cuticular wax and stomatal factor might be involved in regulating water loss in mulberry leaves under field conditions. The variability in moisture retention capacity and cuticular wax characteristics might be important in evaluating and screening mulberry cultivars for increasing silk quality and silkworm productivity.     

Fine mapping of BrWax1, a gene controlling cuticular wax biosynthesis in Chinese cabbage (Brassica rapa L. ssp. pekinensis)

The surface of plants is covered with a cuticular wax, which contains a mixture of very-long-chain fatty acid derivatives. This wax layer provides a hydrophobic barrier which reduces non-stomatal water loss and prevents pathogen attack. The biosynthesis pathway of cuticular wax in Arabidopsis is well studied; however, little is known about the synthesis of cuticular wax in Brassica rapa. Genetic analyses indicated that the waxy characteristic is controlled by a single dominant gene. In the present study, preliminary mapping results from an F2 population consisting of 308 recessive individuals showed that the BrWax1 (Brassica Wax) gene is located on linkage group A01. We developed a set of new markers closely linked to the target gene, and used another population of 1,020 recessive F2 individuals to fine-map the BrWax1 gene to a genomic DNA fragment of approximately 86.4 kb. Fifteen genes were identified in this target region. Based on gene annotation, the Bra013809 gene was the candidate for the BrWax1 gene. Quantitative real-time PCR analysis and expression pattern of the two parents showed that the expression level of Bra013809 was much higher in the waxy phenotype than in the glossy phenotype. This result should provide not only important information for functional studies of the BrWax1 gene, but also the starting point for studying the pathway of cuticular wax biosynthesis in Brassica rapa.     

OsGL1-3 is Involved in Cuticular Wax Biosynthesis and Tolerance to Water Deficit in Rice

Cuticular wax covers aerial organs of plants and functions as the outermost barrier against non-stomatal water loss. We reported here the functional characterization of the Glossy1(GL1)-homologous gene OsGL1-3 in rice using overexpression and RNAi transgenic rice plants. OsGL1-3 gene was ubiquitously expressed at different level in rice plants except root and its expression was up-regulated under ABA and PEG treatments. The transient expression of OsGL1-3–GFP fusion protein indicated that OsGL1-3 is mainly localized in the plasma membrane. Compared to the wild type, overexpression rice plants exhibited stunted growth, more wax crystallization on leaf surface, and significantly increased total cuticular wax load due to the prominent changes of C₃₀–C₃₂ aldehydes and C₃₀ primary alcohols. While the RNAi knockdown mutant of OsGL1-3 exhibited no significant difference in plant height, but less wax crystallization and decreased total cuticular wax accumulation on leaf surface. All these evidences, together with the effects of OsGL1-3 on the expression of some wax synthesis related genes, suggest that OsGL1-3 is involved in cuticular wax biosynthesis. Overexpression of OsGL1-3 decreased chlorophyll leaching and water loss rate whereas increased tolerance to water deficit at both seedling and late-tillering stages, suggesting an important role of OsGL1-3 in drought tolerance.     

Leaf cuticular waxes and physiological parameters in alfalfa leaves as influenced by drought

Drought significantly constrains higher yield of alfalfa (Medicago sativa L.) in arid and semiarid areas all over the world. This study evaluated the responses of leaf cuticular wax constituents to drought treatment and their relations to gas-exchange indexes across six alfalfa cultivars widely grown in China. Water deficit was imposed by withholding water for 12 d during branching stage. Cuticular waxes on alfalfa leaves were dominated by primary alcohols (41.7?C54.2%), alkanes (13.2?C26.9%) and terpenes (17.5?C28.9%), with small amount of aldehydes (1.4?C3.4%) and unknown constituents (4.5?C18.4%). Compared to total wax contents, the wax constituents were more sensitive to drought treatment. Drought decreased the contents of primary alcohol and increased alkanes in all cultivars. Alkane homologs, C25, C27, and C29, were all negatively correlated with photosynthetic rate, transpiration rate, stomatal conductance, and leaf water potential. Under drought conditions, both stomatal and nonstomatal factors were involved in controlling water loss from alfalfa leaves. No direct relationship was observed between wax contents and drought resistance among alfalfa cultivars. An increase in alkane content might be more important in improving drought tolerance of alfalfa under water deficit, which might be used as an index for selecting and breeding drought resistant cultivars of alfalfa.     

Fine mapping of <Emphasis Type="Italic">BoGL1</Emphasis>, a gene controlling the glossy green trait in cabbage (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. Var. <Emphasis Type="Italic">capitata</Emphasis>)

The cuticular wax covering epidermal cells causes the glaucous appearance in cabbage. As a protective barrier, cuticular wax plays various roles in protection against biotic and abiotic stresses. This is the first gene mapping report of a dominant glossy green cabbage mutant. In the present paper, scanning electron microscopy (SEM) demonstrated that the wax crystals were severely reduced in the mutant, which indicates that the glossy green phenotype is caused by cuticular wax reduction. Genetic analysis revealed that the glossy trait is controlled by a single dominant gene. Through primer screening and fine mapping, the mutant gene BoGL1 (Brassica oleracea glossy 1) was delimited to the end of chromosome C08 by the flanking marker SSRC08–76 at a genetic distance of 0.2 cM. Two genes homologous to CER1 (ECERIFERUM 1), a gene related to wax biosynthesis in Arabidopsis, were located in the mapped region. Expressional analysis revealed that the Bol018504 gene was severely suppressed but that no nucleotide variation was found by sequencing. These results lay the foundation for the functional analysis of BoGL1, and they will accelerate the research on wax metabolism in cabbage.     

Crystal structures and chemical composition of leaf surface wax depositions on the desert moss Syntrichia caninervis

The structures and composition of cuticular waxes deposited on the leaves of a typical desert moss, Syntrichia caninervis, were investigated. The wax crystals deposited on leaves shifted with leaf aging. The results of chemical analysis showed the main chemical components of the moss wax were fatty acids, alcohols and alkanes. Leaf aging increased the content of cuticular wax and the percentage of very long chain components, from 1150 μg g⁻¹ DW and 13.6% in younger leaves to 2640 μg g⁻¹ DW and 37.2% in aged leaves, respectively. Dehydration/hydration also augmented the wax content by 35.17% in juvenile leaves and by 1900% in lab-cultivated leaves after three-cycle treatments. Synthesis of hexadecanoic acid and tetracosane were predicted to be the first step of wax accumulation. The responses of cuticular waxes in crystal structure and chemical composition were recommended as a biomonitor for assessing the shift of ecological and environmental quality.     

Cuticular wax variation in the tomato (Solanum lycopersicum L.), related wild species and their interspecific hybrids

The tomato (Solanum lycopersicum L.) is one of the world's most important vegetable crop species. Among the many tomato accessions available, only a few are tolerant to abiotic stresses, which are responsible for the majority of the crop losses worldwide. Wild tomato species are then secondary gene pool in the breeding of more resistant tomato cultivars. In the current study, the composition of leaf cuticular waxes from fourteen tomato accessions, including S. lycopersicum, Solanum pennellii, Solanum pimpinellifolium, and their interspecific hybrids was studied in order to select the most adequate chemotaxonomic markers. Total cuticular wax load of S. pennellii plants was much higher than in the other plant species. Hydrocarbons were usually the most abundant wax components, followed by minor quantities of triterpenes and other compounds. Interspecific hybrids showed intermediate wax characteristics. The amount and composition of surface waxes were not correlated with the abiotic stress tolerance in S. lycopersicum. The composition of the hydrocarbon fraction was the least variable both within a single accession and between all the plants studied. Based on the results, cuticular hydrocarbons are proposed as potential chemotaxonomic markers in the classification of tomato and related species.