Plant Cuticular Waxes Research Articles

Soil pH is the Primary Factor Driving the Transformation of Plant Cuticular Wax in Saline Soils

Soil pH is the Primary Factor Driving the Transformation of Plant Cuticular Wax in Saline Soils

Chemical and Transcriptomic Analyses of Leaf Cuticular Wax Metabolism in Ammopiptanthus mongolicus under Osmotic Stress.

Plant cuticular wax forms a hydrophobic structure in the cuticle layer covering epidermis as the first barrier between plants and environments. Ammopiptanthus mongolicus, a leguminous desert shrub, exhibits high tolerances to multiple abiotic stress. The physiological, chemical, and transcriptomic analyses of epidermal permeability, cuticular wax metabolism and related gene expression profiles under osmotic stress in A. mongolicus leaves were performed. Physiological analyses revealed decreased leaf epidermal permeability under osmotic stress. Chemical analyses revealed saturated straight-chain alkanes as major components of leaf cuticular wax, and under osmotic stress, the contents of total wax and multiple alkane components significantly increased. Transcriptome analyses revealed the up-regulation of genes involved in biosynthesis of very-long-chain fatty acids and alkanes and wax transportation under osmotic stress. Weighted gene co-expression network analysis identified 17 modules and 6 hub genes related to wax accumulation, including 5 enzyme genes coding KCS, KCR, WAX2, FAR, and LACS, and an ABCG transporter gene. Our findings indicated that the leaf epidermal permeability of A. mongolicus decreased under osmotic stress to inhibit water loss via regulating the expression of wax-related enzyme and transporter genes, further promoting cuticular wax accumulation. This study provided new evidence for understanding the roles of cuticle lipids in abiotic stress tolerance of desert plants.

The cuticular wax on sorghum straw influenced soil microbial diversity and straw degradation in soil

The cuticular waxes cover living plant surface but the ecological significance of cuticular wax in crop straw returning to soil remains uncertain. This study focused on sorghum straw to investigate the bioactivity of cuticular waxes in the soil system. The surface of sorghum leaf, sheath and stem organs was covered with epicuticular wax crystals, which present a dull and glaucous wax layer. The leaf surface with wax exhibited an average contact angle of 83°, while after the wax removal the contact angle decreased to 53° on average, showing hydrophobic surface on the sorghum leaf. In total, six wax classes were identified across all organs, including aldehydes, alkanes, fatty acids, primary alcohols, sterols, and minor triterpenoids. The total wax coverage on leaves was 1.5 μg·cm−2, which was significantly lower than that on sheaths (20.6 μg·cm−2) and stems (6.3 μg·cm−2). The wax addition increased the proportions of Actinobacteria and Firmicutes by 24.42 % and 78.02 %, whereas decreased Proteobacteria and Acidobacteria by 16.32 % and 23.16 % on day 14 under higher wax rate. The microbial communities inhabiting returned straw surfaces also differed significantly between wax-retrained and wax-removed straw, as well as among leaf, stem, and sheath components, and at various decomposition stages. Wax addition increased soil microbial biomass carbon and nitrogen by 56.76 % and 25.60 % at higher rate, and soil enzyme activities (i.e., soil urease, soil acid phosphatase activity, soil sucrase activity). Wax removal significantly increased the decomposition rate of dry matter by 24.28 % and crude fiber by 27.22 % on day 154. These findings demonstrate that plant cuticular waxes maintain their bioactivities after entering the soil, which broadens our knowledge of the roles of plant cuticular waxes in ecosystems. Therefore, selecting efficient wax-degrading microorganisms would accelerate the nutrient cycling of crops in soil system.

Mapping of the Waxy Gene in Brassica napus L. via Bulked Segregant Analysis (BSA) and Whole-Genome Resequencing

Plant cuticular wax is the covering of the outer layer of the plant. It forms a protective barrier on the epidermis of plants and plays a vital role like a safeguard from abiotic and biotic stresses. In the present study, Brassica napus L. materials with and without wax powder were observed. Genetic analysis showed that the separation ratio of waxy plants to waxless plants was 15:1 in the F2 population, which indicated that the wax powder formation was controlled by two pairs of genes. In order to identify the candidate genes associated with the wax powder trait of B. napus L., bulked segregant analysis (BSA) was performed. The homozygous waxy plants, the homozygous waxless plants, and plants from three parents were selected for establishing five DNA pools for genome-wide resequencing. The results of the resequencing showed that the site associated with wax powder trait was located in the region of 590,663–1,657,546 bp on chromosome A08. And 48 single nucleotide polymorphisms (SNPs) were found between the DNA sequences of waxy plants and waxless plants in this region. These SNPs were distributed across 16 gene loci. qRT-PCR analysis was conducted for the 16 candidate genes and three genes (BnaA08g01070D, BnaA08g02130D, and BnaA08g00890D) showed significantly differential expression between waxy and waxless parents. BnaA08g01070D and BnaA08g02130D were significantly down-regulated in the waxless parent, while BnaA08g00890D was significantly up-regulated in the waxless parent. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed that the BnaA08g02130D gene was enriched in lipid biosynthetic or metabolic processes. All the results in our study would provide valuable clues for exploring the genes involved in wax powder development.

Eceriferum Genes in Tomato (Solanum lycopersicum): Genome-Wide Identification and Expression Analysis Reveal Their Potential Functions during Domestication

Plant cuticular wax plays an important role in resistance to environmental stresses. Eceriferum (CER) genes are involved in wax synthesis. However, little information is available for tomato species. In this study, 26 SlCER genes were identified in tomato (S. lycopersicum), and they were classified into four clades. The physicochemical properties and conserved motifs of their proteins were predicted. These SlCERs were mainly expressed in leaves, flowers or fruits, and most SlCERs played roles in response to abiotic stresses, especially drought stress. Furthermore, the changes in haplotypes indicated that SlCERs might have been involved in adapting to the environments for wild species S. pimpinellifolium before domestication. These findings would lay a foundation for future functional studies of SlCERs and also provide insights for anti-stress improvement in tomato in the near future.

An ancestral role for 3-KETOACYL-COA SYNTHASE3 as a negative regulator of plant cuticular wax synthesis.

The plant cuticle, a structure primarily composed of wax and cutin, forms a continuous coating over most aerial plant surfaces. The cuticle plays important roles in plant tolerance to environmental stress, including stress imposed by drought. Some members of the 3-KETOACYL-COA SYNTHASE (KCS) family are known to act as metabolic enzymes involved in cuticular wax production. Here we report that Arabidopsis (Arabidopsis thaliana) KCS3, which was previously shown to lack canonical catalytic activity, instead functions as a negative regulator of wax metabolism by reducing the enzymatic activity of KCS6, a key KCS involved in wax production. We demonstrate that the role of KCS3 in regulating KCS6 activity involves physical interactions between specific subunits of the fatty acid elongation complex and is essential for maintaining wax homeostasis. We also show that the role of the KCS3-KCS6 module in regulating wax synthesis is highly conserved across diverse plant taxa from Arabidopsis to the moss Physcomitrium patens, pointing to a critical ancient and basal function of this module in finely regulating wax synthesis.

Rotator Phases of Aliphatic Aldehydes and Implications for Wax Crystal Growth in Plants

The even C22–32 aliphatic aldehydes were synthesized and analyzed by differential scanning calorimetry and high-temperature X-ray diffraction. Aldehydes were determined to crystallize into triclinic, monoclinic, and orthorhombic lattices and undergo solid–solid transitions into the rotator (R) phase. All of the known R phases for the even and odd n-alkanes were observed for the even aldehydes. Further, binary mixtures of triacontanal and n-nonacosane, two representative constituents of plant cuticular waxes, were determined to be partially miscible at three compositions (25:75, 50:50, 75:25) and likewise possess R-type phases. Many plants exhibit mutations wherein the expression of aldehydes in cuticular waxes is significantly decreased. Afflicted mutants are often glabrous, devoid of epicuticular wax crystals, and suffer from increased cuticular permeability as well as drought intolerance. We propose that aldehydes serve as miscibility intermediaries and their R phases may provide a mechanism for wax crystal growth in plants.

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.

Genome-wide identification of the CER1 gene family in apple and response of MdCER1-1 to drought stress.

Plant cuticular wax was a major consideration affecting the growth and quality of plants through protecting the plant from drought and other diseases. According to existing studies, CER1, as the core enzyme encoding the synthesis of alkanes, the main component of wax, can directly affect the response of plants to stress. However, there were few studies on the related functions of CER1 in apple. In this study, three MdCER1 genes in Malus domestica were identified and named MdCER1-1, MdCER1-2, and MdCER1-3 according to their distribution on chromosomes. Then, their physicochemical properties, sequence characteristics, and expression patterns were analyzed. MdCER1-1, with the highest expression level among the three members, was screened for cloning and functional verification. Real-time fluorescence quantitative PCR (qRT-PCR) analysis also showed that drought stress could increase the expression level of MdCER1-1. The experiment of water loss showed that overexpression of MdCER1-1 could effectively prevent water loss in apple calli, and the effect was more significant under drought stress. Meanwhile, MdYPB5, MdCER3, and MdKCS1 were significantly up-regulated, which would be bound up with waxy metabolism. Gas chromatography-mass spectrometer assay of wax fraction makes known that overexpression of MdCER1-1 apparently scaled up capacity of alkanes. The enzyme activities (SOD, POD) of overexpressed apple calli increased significantly, while the contents of proline increased compared with wild-type calli. In conclusion, MdCER1-1 can resist drought stress by reducing water loss in apple calli epidermis, increasing alkanes component content, stimulating the expression of waxy related genes (MdYPB5, MdCER3, and MdKCS1), and increasing antioxidant enzyme activity, which also provided a theoretical basis for exploring the role of waxy in other stresses.

Two Arabidopsis MYB-SHAQKYF transcription repressors regulate leaf wax biosynthesis via transcriptional suppression on DEWAX.

Plant cuticular wax accumulation limits nonstomatal transpiration and is regulated by external environmental stresses. DEWAX (DECREASE WAX BIOSYNTHESIS) plays a vital role in diurnal wax biosynthesis. However, how DEWAX expression is controlled and the molecular mechanism of wax biosynthesis regulated by the diurnal cycle remains largely unknown. Here, we identified two Arabidopsis MYB-SHAQKYF transcription factors, MYS1 and MYS2, as new regulators in wax biosynthesis and drought tolerance. Mutations of both MYS1 and MYS2 caused significantly reduced leaf wax, whereas overexpression of MYS1 or MYS2 increased leaf wax biosynthesis and enhanced drought tolerance. Our results demonstrated that MYS1 and MYS2 act as transcription repressors and directly suppress DEWAX expression via ethylene response factor-associated amphiphilic repression motifs. Genetic interaction analysis with DEWAX, SPL9 (SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 9), and CER1 (ECERIFERUM 1) in wax biosynthesis and under drought stresses demonstrated that MYS1 and MYS2 act upstream of the DEWAX-SPL9 module, thus regulating CER1 expression. Expression analysis suggested that the diurnal expression pattern of DEWAX is partly regulated by MYS1 and MYS2. Our findings demonstrate the roles of two unidentified transcription repressors, MYS1 and MYS2, in wax biosynthesis and provide insights into the mechanism of diurnal cycle-regulated wax biosynthesis.

Evolution and molecular basis of substrate specificity in a 3-ketoacyl-CoA synthase gene cluster from Populus trichocarpa

Very long chain fatty acids (VLCFAs) are precursors to sphingolipids, glycerophospholipids, and plant cuticular waxes. In plants, members of a large 3-ketoacyl-CoA synthase (KCS) gene family catalyze the substrate-specific elongation of VLCFAs. Although it is well understood that KCSs have evolved to use diverse substrates, the underlying molecular determinants of their specificity are still unclear. In this study, we exploited the sequence similarity of a KCS gene cluster from Populus trichocarpa to examine the evolution and molecular determinants of KCS substrate specificity. Functional characterization of five members (PtKCS1, 2, 4, 8, 9) in yeast showed divergent product profiles based on VLCFA length, saturation, and position of the double bond. In addition, homology models, rationally designed chimeras, and site-directed mutants were used to identify two key regions (helix-4 and position 277) as being major determinants of substrate specificity. These results were corroborated with chimeras involving a more distantly related KCS, PtCER6 (the poplar ortholog of the Arabidopsis CER6), and used to show that helix-4 is necessary for the modulatory effect of PtCER2-like5 on KCS substrate specificity. The role of position 277 in limiting product length was further tested by substitution with smaller amino acids, which shifted specificity toward longer products. Finally, treatment with KCS inhibitors (K3 herbicides) showed varying inhibitor sensitivities between the duplicated paralogs despite their sequence similarity. Together, this work sheds light on the molecular mechanisms driving substrate diversification in the KCS family and lays the groundwork for tailoring the production of specific VLCFAs.

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Context Plant species exhibit different patterns of plant cuticular wax profiles, which can potentially be used as diet composition markers in free-grazing herbivores. Aims Evaluate the suitability of the plant cuticular n-alkanes, long-chain alcohol (LCOH) profiles and carbon stable isotope enrichment (δ13C) of n-alkanes as markers in the estimation of diet composition of grazing animals. Methods Forage samples were collected from 100 representative quadrats of 0.5 m × 0.5 m at 10 m transects and sorted by species and pooled from different quadrats to obtain enough quantities of representative individual species. In total, 10 dominant forage species were identified and analysed for n-alkanes and LCOH by gas chromatography, and the isotopic ratio by using gas chromatography–combustion–isotope ratio mass spectrometry. Principal component analysis (PCA) was used to identify inter-species differences in the concentration patterns of plant wax components. Key results Odd-chain n-alkanes comprised the highest proportion of the total n-alkane concentration, ranging from 79% in Ischaemum afrum to 95% in Haplocarpha hastata. n-Alkanes C31, C29 and C33 were the most abundant with an average of 167, 80 and 61 mg/kg DM, in that order, in all species. Even-chain LCOH comprised the highest proportion of the total LCOH concentration, accounting for 92% in Brachiaria scalaris to 97% in Ischaemum afrum. The dominant even-chain LCOH were C30OH, C32OH, C28OH and C26OH, with an average concentration of 362, 348, 266 and 237 mg/kg DM respectively, across species. The δ13C of n-alkanes showed large variations among forage species, ranging from −19.7‰ in Andropogon amethystinus to −40.6‰ in Trifolium mattirolianum. The result of the PCA showed that 81% of the variance in the pattern of concentrations of n-alkanes was explained by the first two principal components compared with 69.3% and 82.9% in the case of LCOH and δ13C of n-alkanes, respectively. Conclusions Noticeable variations were observed for forage species studied in the patterns of plant wax components. Implications The differences in the patterns of concentrations of n-alkanes, LCOH and δ13C of n-alkanes could be suitable as markers for diet composition estimation of grazing animals.

Environment-Driven Adaptations of Leaf Cuticular Waxes Are Inheritable for Medicago ruthenica.

Cuticular waxes covering the plant surface play pivotal roles in helping plants adapt to changing environments. However, it is still not clear whether the responses of plant cuticular waxes to their growing environments are inheritable. We collected seeds of Medicago ruthenica (a perennial legume) populations from 30 growing sites in northern China and examined the variations of leaf cuticular waxes in a common garden experiment. Four wax genes, MrFAR3-1, MrFAR3-2, MrCER1, and MrKCS1, involved in biosynthesis of predominant wax classes (primary alcohol and alkane) and wax precursors, were isolated to test the contributions of genetic variations of the coding sequences (CDS) and the promoter sequences and epigenetic modifications. The plasticity responses of the cuticular waxes were further validated by two stress-modeling experiments (drought and enhancing ultraviolet B). Great variations in total wax coverage and abundance of wax classes or wax compounds were observed among M. ruthenica populations in a common garden experiment. Stress-modeling experiments further validated that M. ruthenica would alter leaf wax depositions under changed growing conditions. The transcriptional levels of the wax genes were positively or negatively correlated with amounts of cuticular waxes. However, the analysis of promoter methylation showed that the methylation level of the promoter region was not associated with their expressions. Although both promoter sequences and CDS showed a number of polymorphic sites, the promoters were not naturally selected and insignificant difference could be observed in the numbers and types of acting elements of the four wax genes among populations. In contrast, the CDS of the wax genes were naturally selected, with a number of missense mutations resulting in alterations of the amino acid as well as their isoelectric points and polarities, which could impact on enzyme function/activity. We conclude that long-term adaptation under certain environments would induce genetic mutation of wax biosynthesis genes, resulting in inheritable alterations of cuticular wax depositions.

Regulation of cuticular wax biosynthesis in plants under abiotic stress

Cuticular waxes are the covering of the outer layer of the plant, consist of hydrocarbon appears like whitish film or bloom in plant organs. They play a vital role like a safeguard from different stress condition in the plant. Since environmental factors are active regulators of cuticular wax biosynthesis, composition, quantity, and deposition, it is evident that cuticular wax is associated with plant stress responses. The diversity of cuticular wax compositions is a proof of the wealth of genes associated in plant wax production. Moreover, a number of wax genes were distinguished in plant/crops at abiotic stress conditions but, regulation of control of those wax genes has not been studied very well in major crop plants at abiotic conditions. A very few transcriptions factors were identified to regulate the expression level of wax genes of cuticular wax biosynthesis at abiotic stress condition. However, further study is needed to identify more candidate transcriptional regulation factors to cuticular wax production in different crop plants in diverse abiotic environments. Therefore, regulation of cuticular wax production under diverse abiotic stresses and the role of transcription factors into the plant cuticular wax accumulation will be helpful to engineer crop plants and improve transgenic crops for stress tolerance. In this review, we focused on a new perspective on transcriptional factors to regulate functional genes of cuticular wax biosynthesis in plants at abiotic stresses.

Progress of Plant Cuticular Wax Synthesis and Its Regulatory Factor WIN/SHN

Progress of Plant Cuticular Wax Synthesis and Its Regulatory Factor WIN/SHN

The chemical composition and potential role of epicuticular and intracuticular wax in four cultivars of table grapes

Abstract Plant cuticular wax is the first barrier to resist biotic and abiotic stress. However, little is known about the compositional differences between the epicuticular and intracuticular wax in grape berry. The compositional, morphological and functional features of cuticular wax in grape berries of Vitis vinifera cv. ‘Kyoho’, ‘Muscat Hamburg’, ‘Redglobe’, and ‘Zuijinxiang’ were investigated. The total and epicuticular wax of four berries were mainly composed of terpenoids, hydrocarbons, alcohols, fatty acids and esters. Oleanolic acid was the most abundant terpenoid among the four cultivars. Scanning electron microscopy revealed the crystalline flakes structure of the cuticular wax. Additionally, the removal of epicuticular wax accelerated the weight loss, browning, and softening of grape berries, indicating the plastic-wrap-like effect of the cuticular wax on postharvest quality. This study provided the theoretical basis for further application of the fruit wax or waxy analogue.

Update on Cuticular Wax Biosynthesis and Its Roles in Plant Disease Resistance.

The aerial surface of higher plants is covered by a hydrophobic layer of cuticular waxes to protect plant tissues against enormous environmental challenges including the infection of various pathogens. As the first contact site between plants and pathogens, the layer of cuticular waxes could function as a plant physical barrier that limits the entry of pathogens, acts as a reservoir of signals to trigger plant defense responses, and even gives cues exploited by pathogens to initiate their infection processes. Past decades have seen unprecedented proceedings in understanding the molecular mechanisms underlying the biosynthesis of plant cuticular waxes and their functions regulating plant–pathogen interactions. In this review, we summarized the recent progress in the molecular biology of cuticular wax biosynthesis and highlighted its multiple roles in plant disease resistance against bacterial, fungal, and insect pathogens.

Sequestration of Exogenous Volatiles by Plant Cuticular Waxes as a Mechanism of Passive Associational Resistance: A Proof of Concept.

Numerous plant-derived volatile organic compounds (VOCs) induce the expression of resistance-related genes and thereby cause an “associational resistance” in neighbouring plants. However, VOCs can also be sequestered by plant cuticular waxes. In case that they maintain their biological activity, such sequestered VOCs could generate a “passive” associational resistance that is independent of any gene expression in the receiver. As a proof of concept, we used major components of the cuticular wax layers of the tree, Parkinsonia praecox, and conidia of Colletotrichum lindemuthianum, a fungal pathogen that has not been reported to infect P. praecox. Wax layers were re-constituted on glass slides and exposed to each of 20 pure VOCs for 1 d and then to ambient air for 1 d or 15 d. Gas chromatography-mass spectrometry (GC-MS) analyses showed that all 20 VOCs were sequestered by the re-constituted wax layers. Exposure to 18 of the VOCs significantly inhibited the germination of C. lindemuthianum conidia on these wax layers after 1 day of exposure to ambient air. Four of the VOCs: 4Z-heptenol, farnesene, limonene, and 2E-decenal, inhibited germination rates to less than 25% of the controls. After 15 d, all VOCs were still detectable, although at strongly reduced concentrations, and no significant inhibition of conidial germination could be detected anymore. Exogenous VOCs can be sequestered by the components of plant cuticular waxes and maintain their biological activity, at least over a certain time span: an effect that could generate a transient “passive associational resistance” to pathogens.

Cytochrome P450 family member CYP96B5 hydroxylates alkanes to primary alcohols and is involved in rice leaf cuticular wax synthesis.

Odd-numbered primary alcohols are components of plant cuticular wax, but their biosynthesis remains unknown. We isolated a rice wax crystal-sparse leaf5 (WSL5) gene using a map-based cloning strategy. The function of WSL5 was illustrated by overexpression and knockout in rice, heterologous expression in Arabidopsis and transient expression in tobacco leaves. WSL5 is predicted to encode a cytochrome P450 family member CYP96B5. The wsl5 mutant lacked crystalloid platelets on the surface of cuticle membrane, and its cuticle membrane was thicker than that of the wild-type. The wsl5 mutant is more tolerant to drought stress. The load of C23 -C33 alkanes increased, whereas the C29 primary alcohol reduced significantly in wsl5 mutant and WSL5 knockout transgenic plants. Overexpression of WSL5 increased the C29 primary alcohol and decreased alkanes in rice leaves. Heterologous expression of WSL5 increased the C29 primary alcohol and decreased alkanes, secondary alcohol, and ketone in Arabidopsis stem wax. Transient expression of WSL5 in tobacco leaves also increased the production C29 primary alcohol. WSL5 catalyzes the terminal hydroxylation of alkanes, yielding odd-numbered primary alcohols, and is involved in the formation of epidermal wax crystals on rice leaf, affecting drought sensitivity.

Variations of soil organic matters and plant cuticular waxes along an altitude gradient in Qinghai-Tibet Plateau

This study aims to evaluate the variations of soil organic matter and plant leaf cuticular waxes along an altitude gradient at two mountains in the east side of the Qinghai-Tibet Plateau. Surface soils were sampled along an altitude gradient (2600, 3000, 3400, 3800 and 4200 m). We analyzed the soil total organic carbon (TOC), dissolved organic carbon (DOC), soil lipids and leaf cuticular waxes in Kobresia humilis which distributed at all tested altitudes. A sharp concentration increase of TOC was observed when altitudes increased, whereas the proportions of DOC in TOC at low altitudes (3000 m and 2600 m) were significantly higher than those at higher altitudes. Altitudes also influenced the molecular weight, humification degree, aromaticity, and hydrophobicity of DOC molecular. Though soils at higher altitudes relatively had higher abundance of odd carbon alkanes and even carbon alcohols, soil lipid concentrations showed no significant difference between altitudes. Relatively higher total wax coverage at high and low altitude was observed on leaves of K. humilis. The changes of altitudes at alpine ecosystem were associated with alterations of both quantity and quality of soil organic matter as well as leaf cuticular wax coverage, which contributed to their environmental adaptations.