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PubMed2001

Identification of chemical components of corn kernel pericarp wax associated with resistance to Aspergillus flavus infection and aflatoxin production.

Kernel pericarp wax of the corn breeding population GT-MAS:gk has been associated with resistance to Aspergillus flavus infection and aflatoxin production. GT-MAS:gk wax, previously compared to waxes of three susceptible genotypes, was presently compared to wax of a different, and more numerous, group of susceptible lines. Wax separation by TLC confirmed previous findings, demonstrating a unique GT-MAS:gk band and a unique "susceptible" band. Only GT-MAS:gk wax inhibited the growth of A. flavus; however, no association was established, as before, between kernel wax abundance and resistance. Gas chromatography-mass spectroscopy (GC-MS) analysis of kernel whole wax showed a higher percentage of phenol-like compounds in wax from GT-MAS:gk than in waxes from the susceptible lines. The GT-MAS:gk unique band contained phenol-like compounds and ethyl-hexadecanoate; butyl-hexadecanoate was preeminent in most of the "susceptible bands". Alkylresorcinol (phenolic compounds) content was dramatically higher in GT-MAS:gk wax than in the wax of susceptible lines. An alkylresorcinol, 5-methylresorcinol, also inhibited in vitro growth of A. flavus. These and other phenolic compounds may contribute to kernel wax inhibition of A. flavus infection/aflatoxin production. Further investigation is needed to confirm a role for them in GT-MAS:gk resistance.

Aflatoxins↗
PubMed2002

Efficacy of a wax containing benzocaine in the relief of oral mucosal pain caused by orthodontic appliances.

During orthodontic treatment, pain and discomfort of the oral mucosa can be experienced as a result of trauma from the appliances caused by increased friction between mucosal tissue and the surface of the brackets. Currently, orthodontists have few remedies to prevent or relieve this mucosal irritation. The orthodontist can give the patient wax to cover the brackets as a prophylactic measure or to use as needed on specific irritating appliances. Orthodontic wax contains no analgesic components. The purpose of this study was to determine the efficacy of an orthodontic wax containing benzocaine that is released over time in a controlled manner. This randomized, prospective, double-blind, clinical trial compared patients' responses to the wax with benzocaine and the currently used unmedicated orthodontic wax. Seventy patients, 35 in each treatment group, were instructed to apply the wax in a specific manner, and their pain levels were recorded at 6 different time points. The pain levels were analyzed with a repeated-measure analysis of variance (ANOVA) model with factors of treatment (medicated wax vs unmedicated wax) and times (6 levels). Post-hoc pair-wise comparisons were made on the basis of the Fisher least significant difference procedure. The results of this study clearly indicated that the pain profile over time of the group that received the wax containing benzocaine was significantly different from that of the subjects who received the unmedicated wax. The medicated group had significantly lower pain levels at every time point after the first hour (P <.0003 in each case) compared with the unmedicated group. The medicated wax was effective immediately and continued to reduce pain in greater magnitude than did the unmedicated wax. A wax applied to orthodontic brackets that slowly and continuously releases benzocaine is significantly more effective at reducing the pain associated with mucosal irritation than is the current option used by most orthodontists.

Adult↗
PubMed2003

Molecular and carbon isotopic composition of leaf wax in vegetation and aerosols in a northern prairie ecosystem.

We measured the molecular and carbon isotopic composition of major leaf wax compound classes in northern mixed mesic prairie species (Agropyron smithii, Stipa viridula, Bouteloua gracilis, Tragopogon dubius) and in selected crops (Triticum aestivum, Brassica napus, Hordeum vulgare, Medicago sativa) of southern Alberta and also in aerosols collected 4 m above the prairie canopy. Our aims were to better constrain the wax biosynthetic carbon isotopic fractionation relative to the plant's carbon isotopic discrimination and to quantitatively assess the correspondence between wax composition in vegetation and in boundary layer aerosols. Wax molecular composition of the C(3)prairie species and bulked vegetation was characterized by high abundance of C(28) n-alkanol and C(31) n-alkane compounds whereas the C(4) species B. gracilis had several co-dominant n-alkanol and n-alkane compounds. Wax molecular composition of crop species differed significantly from that of prairie vegetation and was often dominated by a single compound. Results indicate that leaf wax isotopic composition is quantitatively related to the plant's carbon isotopic discrimination. Although species variations were evident, n-alcohol, n-acid and n-alkane wax compounds were on average depleted in (13)C by approximately 6.0+/-1 per thousand relative to total plant carbon. The magnitude of the depletion in wax delta(13)C was unaffected by environmental factors which altered photosynthetic carbon isotopic discrimination. No consistent difference in the magnitude of wax biosynthetic fractionation was observed between C(3) and C(4) species, indicating that photosynthetic pathway has little influence on the isotopic fractionation of wax during biosynthesis. The isotopic composition of ablated waxes in aerosols collected above the canopy was similar to that of the grassland vegetation but the molecular composition differed significantly and indicated that the source "footprint" of the ablated leaf wax particles we sampled in boundary layer air masses was of a regional or larger spatial scale.

Aerosols↗
PubMed2002

Diversity of cuticular wax among Salix species and Populus species hybrids.

The leaf cuticular waxes of three Salix species and two Populus species hybrids, selected for their ability to produce high amounts of biomass, were characterized. Samples were extracted in CH(2)Cl(2) three times over the growing season. Low kV SEM was utilized to observe differences in the ultrastructure of leaf surfaces from each clone. Homologous series of wax components were classified into organic groups, and the variation in wax components due to clone, sample time, and their interaction was identified. All Salix species and Populus species hybrids showed differences in total wax load at each sampling period, whereas the pattern of wax deposition over time differed only between the Salix species. A strong positive relationship was identified between the entire homologous series of alcohols and total wax load in all clones. Similarly strong relationships were observed between fatty acids and total wax load as well as fatty acids and alcohols in two Salix species and one Populus species hybrid. One Salix species, S. dasyclados, also displayed a strong positive relationship between alcohols and alkanes. These data indicate that species grown under the same environmental conditions produce measurably different cuticular waxes and that regulation of wax production appears to be different in each species. The important roles cuticular waxes play in drought tolerance, pest, and pathogen resistance, as well as the ease of wax extraction and analysis, strongly suggest that the characteristics of the cuticular wax may prove to be useful selectable traits in a breeding program.

Hybridization, Genetic↗
PubMed1999

CUT1, an Arabidopsis gene required for cuticular wax biosynthesis and pollen fertility, encodes a very-long-chain fatty acid condensing enzyme.

Land plants secrete a layer of wax onto their aerial surfaces that is essential for survival in a terrestrial environment. This wax is composed of long-chain, aliphatic hydrocarbons derived from very-long-chain fatty acids (VLCFAs). Using the Arabidopsis expressed sequence tag database, we have identified a gene, designated CUT1, that encodes a VLCFA condensing enzyme required for cuticular wax production. Sense suppression of CUT1 in transgenic Arabidopsis plants results in waxless (eceriferum) stems and siliques as well as conditional male sterility. Scanning electron microscopy revealed that this was a severe waxless phenotype, because stems of CUT1-suppressed plants were completely devoid of wax crystals. Furthermore, chemical analyses of waxless plants demonstrated that the stem wax load was reduced to 6 to 7% of wild-type levels. This value is lower than that reported for any of the known eceriferum mutants. The severe waxless phenotype resulted from the downregulation of both the decarbonylation and acyl reduction wax biosynthetic pathways. This result indicates that CUT1 is involved in the production of VLCFA precursors used for the synthesis of all stem wax components in Arabidopsis. In CUT1-suppressed plants, the C24 chain-length wax components predominate, suggesting that CUT1 is required for elongation of C24 VLCFAs. The unique wax composition of CUT1-suppressed plants together with the fact that the location of CUT1 on the genetic map did not coincide with any of the known ECERIFERUM loci suggest that we have identified a novel gene involved in wax biosynthesis. CUT1 is currently the only known gene with a clearly established function in wax production.

Acetyltransferases↗
PubMed2005

Composite structure of the crystalline epicuticular wax layer of the slippery zone in the pitchers of the carnivorous plant Nepenthes alata and its effect on insect attachment.

The slippery zone situated below the peristome inside pitchers of most carnivorous plants from the genus Nepenthes is covered with a thick layer of epicuticular wax. This slippery zone is reported to play a crucial role in animal trapping and prey retention. In N. alata, the wax coverage consists of two clearly distinguished layers. These layers differ in their structure, chemical composition and mechanical properties, and they reduce the insect attachment in different ways. The lower layer resembles foam, composed of interconnected membraneous platelets protruding from the surface at acute angles. The upper layer consists of densely placed separate irregular platelets, located perpendicular to the subjacent layer. Crystals of the upper layer bear small stalks, directed downwards and providing connections to the lower layer. These morphological distinctions correlate with differences in the chemical composition of waxes. The compound classes of alkanes, aldehydes, primary alcohols, free fatty acids, esters and triterpenoids occurred in extracts from both wax layers, but in different proportions. Chain length distributions in aliphatics were different in extracts from the lower and the upper wax layers. Waxes of the upper and lower layers exhibited different mechanical properties: wax of the lower layer is harder and stiffer than that of the upper layer. Moreover, crystals of the upper layer are brittle and may be easily exfoliated or broken to tiny pieces. Laboratory experiments using tethered insects showed that both wax layers reduce the attachment force of insects. It is assumed that a decrease in insect attachment on the two distinct wax layers is provided by the two different mechanisms: (1) crystals of the upper wax layer contaminate insects' adhesive pads; (2) the lower wax layer leads to a reduction of the real contact area of insects' feet with the plant surface.

Animals↗
PubMed2005

Small plot trials documenting effective mating disruption of oriental fruit moth by using high densities of wax-drop pheromone dispensers.

In 2004 field experiments, we compared the effectiveness of various deployment densities of 0.1-ml paraffin wax drops containing 5% pheromone versus Isomate M-Rosso "rope" dispensers for disruption of Grapholita molesta (Busck). Treatments were evaluated in 0.05-ha (12-tree) plots of 'Delicious' apples receiving regular maintenance according to growers' standards, but not sprayed with insecticides. The application densities of 0.1-ml wax drops were 3 per tree (820/ha), 10 per tree (2,700/ha), 30 per tree (8,200/ha), and 100 per tree (27,300/ha). Wax drops were compared with 3-ml dispensers of pheromone-containing paraffin wax or Isomate M-Rosso ropes at 1.8 per tree (500/ha) and untreated control plots. Treatments were applied before the start of each of three moth generations. Orientational disruption, as measured by inhibition of moth captures in pheromone-baited delta traps, was greatest in plots that received 100 drops per tree (99.2%) and 30 drops per tree (99.4%). More than 55% of tethered, virgin females were mated in control plots after one night of deployment. However, no mating was recorded at the two highest application densities of wax drops where orientational disruption of traps exceeded 99%. Mating ranged from 7 to 20% among the other treatments, including Isomate rope dispensers. G. molesta males were observed closely approaching pheromone dispensers in plots containing ropes and wax drops, documenting competitive attraction between synthetic pheromone sources and feral females. The majority of observed G. molesta males approached within 60 cm of wax drops or pheromone ropes and departed within 20 s by flying upwind. Thirty wax drops per tree yielded higher mating disruption of G. molesta than did Isomate M-Rosso dispensers deployed at the recommended rate of 500/ha (1.8 per tree). Measurement of release rates confirmed behavioral data indicating that paraffin wax dispensers would need to be applied once per G. molesta generation in Michigan. Paraffin wax drops are a promising technology for moth mating disruption. They are cheaper and easier to produce, require less total pheromone per annual application, and produce better mating disruption at appropriate deployment densities compared with Isomate M-Rosso dispensers under high G. molesta population densities. The cost-effectiveness of this approach will require an appropriate mechanized applicator for wax drops.

Animals↗
PubMed1990

Patient preference for waxed or unwaxed dental floss.

The purpose of this study was to discover patient preference for waxed or unwaxed dental floss, and to learn more about individual flossing habits. One hundred patients randomly presenting for routine dental examinations volunteered to sample a brand of similar-appearing waxed and unwaxed dental floss. After flossing an anterior and a posterior contact area with both types, the patients indicated whether they preferred the waxed or unwaxed floss. The patients also answered questions concerning their flossing habits. Waxed floss was found to be preferred after sampling by 79% and unwaxed by 21%. An additional 50 patients sampled a different brand of waxed and unwaxed floss in a similar manner. In this group 78% preferred the waxed and 22% the unwaxed type. A final group of 50 patients compared mint flavored waxed floss with plain waxed floss of the same brand. In this group 56% preferred the mint flavored waxed floss, 24% the plain waxed floss, and 20% had no preference. A combined total of only 29.5% of the patients claimed to floss daily, 53.5% floss irregularly but at least once a week, while 17% do not floss even once a week. Waxed floss was purchased for home use by a combined patient total of 57.5%, unwaxed by 32%, and neither type by 10.5%. An unsubstantiated belief in the superiority of unwaxed floss has persisted to the present time.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1998

Preliminary evaluation of an aqueous wax emulsion for controlled-release coating.

The purpose of this work was to evaluate the use of an aqueous carnauba wax emulsion (Primafresh HS, Johnson Wax) in a spray-coating process. This involved assessing the effectiveness of the wax in sustaining the release of the drug, theophylline. Second, the process by which the drug was released from the wax-coated pellets was modeled. Finally, a method to determine the optimum blend of pellets with different wax thicknesses, in order to yield a zero-order release profile of the drug, was addressed. Nonpareil pellets were loaded with theophylline using a novel powder coating technique. These drug-loaded pellets were then coated with different levels of carnauba wax in a 6-in. diameter Plexiglas fluid bed with a 3.5-in. diameter Wurster partition. Drug release was measured using a spin-filter dissolution device. The study resulted in continuous carnauba wax coatings which showed sustained drug release profile characteristics typical of a barrier-type, diffusion-controlled system. The effect of varying wax thickness on the release profiles was investigated. It was observed that very high wax loadings would be required to achieve long sustained-release times. The diffusion model, developed to predict the release of the drug, showed good agreement with the experimental data. However, the data exhibited an initial lag-time for drug release which could not be predicted a priori based on the wax coating thickness. A method of mixing pellets with different wax thicknesses was proposed as a way to approximate zero-order release.

Emulsions↗
PubMed2003

Submersed culture production of extracellular wax esters by the marine bacterium Fundibacter jadensis.

Fundibacter jadensis strain T9, a marine gram-negative bacterium, was isolated from the intertidal sediment of the German North Sea coast by our group. The cells were able to produce considerable amounts of extracellular wax esters when cultivated with n-alkanes (hexadecane or tetradecane) as a carbon source. The dependence of wax ester production and the composition of the purified wax on different culture conditions (C:N:P ratio and dissolved oxygen tension) were tested. Our results show that wax ester production was not directly growth-linked. The C:N:P ratio had no significant influence on the yield of alkane-free purified wax. The dissolved oxygen tension affected the produced amount of the alkane-free purified wax and the composition of the purified wax; when lower than 2% it decreased the yield of purified wax and led to an altered wax ester composition. Tetradecane as a carbon source enhanced the spectrum of the wax ester composition.

Alkanes↗
PubMed2000

Molecular analysis of intact preen waxes of Calidris canutus (Aves: Scolopacidae) by gas chromatography/mass spectrometry.

The intact preen wax esters of the red knot Calidris canutus were studied with gas chromatography/mass spectrometry (GC/MS) and GC/MS/MS. In this latter technique, transitions from the molecular ion to fragment ions representing the fatty acid moiety of the wax esters were measured, providing additional resolution to the analysis of wax esters. The C21-C32 wax esters are composed of complex mixtures of hundreds of individual isomers. The odd carbon-numbered wax esters are predominantly composed of even carbon-numbered n-alcohols (C14, C16, and C18) esterified predominantly with odd carbon-numbered 2-methyl fatty acids (C7, C9, C11, and C13), resulting in relatively simple distributions. The even carbon-numbered wax esters show a far more complex distribution due to a number of factors: (i) Their n-alcohol moieties are not dominated by even carbon-numbered n-alcohols esterified with odd carbon-numbered 2-methyl fatty acids, but odd and even carbon-numbered n-alcohols participate in approximately equal amounts; (ii) odd carbon-numbered methyl-branched alcohols participate abundantly in these wax ester clusters; and (iii) with increasing molecular weight, various isomers of the 2,6-, 2,8-, and 2,10-dimethyl branched fatty acids also participate in the even carbon-numbered wax esters. The data demonstrate that there is a clear biosynthetic control on the wax ester composition although the reasons for the complex chemistry of the waxes are not yet understood.

Animals↗
PubMed2006

Changes in leaf cuticular waxes of sesame (Sesamum indicum L.) plants exposed to water deficit.

Sesame (Sesamum indicum L.) is one of the most important oilseed crops, having seeds and oil that are highly valued as a traditional health food. The objective of this study was to evaluate leaf cuticular wax constituents across a diverse selection of sesame cultivars, and the responses of these waxes to drought-induced wilting. Water-deficit was imposed on 18 sesame cultivars by withholding irrigation for 15d during the post-flowering stage, and the effect on seed yield and leaf waxes compared with a well-watered control. Leaf cuticular waxes were dominated by alkanes (59% of total wax), with aldehydes being the next-most abundant class. Compared to well-irrigated plants, drought treatment caused an increase in wax amount on most cultivars, with only three cultivars having a notable reduction. When expressed as an average across all cultivars, drought treatment caused a 30% increase in total wax amount, with a 34% increase in total alkanes, a 13% increase in aldehydes, and a 28% increase in the total of unknowns. In all cultivars, the major alkane constituents were the C27, C29, C31, C33, and C35 homologs, whereas the major aldehydes were the C30, C32, and C34 homologs, and drought exposure had only minor effects on the chain length distribution within these and other wax classes. Drought treatments caused a large decrease in seed yield per plant, but did not affect the mean weight of individual seeds, showing that sesame responds to post-flowering drought by reducing seed numbers, but not seed size. Seed yield was inversely correlated with the total wax amount (-0.466*), indicating that drought induction of leaf wax deposition does not contribute directly to seed set. Further studies are needed to elucidate the ecological role for induction of the alkane metabolic pathway by drought in regulating sesame plant survival and seed development in water-limiting environments.

Plant Epidermis↗
PubMed2006

Nanotubules on plant surfaces: chemical composition of epicuticular wax crystals on needles of Taxus baccata L.

Needles of Taxus baccata L. were covered with tubular epicuticular wax crystals varying in diameters (100 and 250 nm) and lengths (300-500 and 500-1000 nm) on the abaxial and adaxial surfaces, respectively. Various sampling protocols were employed to study the chemical composition of the needle waxes on three different levels of spatial resolution. First, a dipping extraction of whole needles yielded the total cuticular wax mixture consisting of very long chain fatty acids (21%), alkanediols (19%), phenyl esters (15%), and secondary alcohols (9%) together with small amounts of aldehydes, primary alcohols, alkanes, alkyl esters, and tocopherols. Second, waxes from both sides of the needle were sampled separately by brushing with CHCl3-soaked fabric glass. Both sides showed very similar qualitative composition, but differed drastically in quantitative aspects, with nonacosan-10-ol (18%) and alkanediols (33%) dominating the abaxial and adaxial waxes, respectively. Third, the epi- and intracuticular wax layers were selectively sampled by a combination of mechanical wax removal and brushing extraction. This provided direct evidence that the tubular wax crystals contained high percentages of nonacosane-4,10-diol and nonacosane-5,10-diol on the abaxial surface, and nonacosan-10-ol on the adaxial surface of the needles. Together with these compounds, relatively large amounts of fatty acids and smaller percentages of aldehydes, primary alcohols, alkyl esters, and alkanes co-crystallized in the epicuticular layer. In comparison, the intracuticular wax consisted of higher portions of cyclic constituents and aliphatics with relatively high polarity. The formation of the tubular crystals is discussed as a spontaneous physico-chemical process, involving the establishment of gradients between the epi- and intracuticular wax layers and local phase separation.

Chromatography, Gas↗
PubMed2002

Survival of Escherichia coli O157:H7 and Salmonella Muenchen on apples as affected by application of commercial fruit waxes.

Survival of Escherichia coli O157:H7, Salmonella Muenchen, and yeasts and molds on apples as affected by application of five commercial apple waxes was investigated. Red Delicious cv. apples at 21 degrees C were spot inoculated with E. coli O157:H7 and S. Muenchen and spray coated with waxes. Apples sprayed with water served as controls. Apples were dried at either 21 or 55 degrees C for 2 min before subjecting to microbiological analysis after storage for 0, 1, 3, 6, and 12 weeks at 2 or 21 degrees C. Drying temperature did not significantly influence populations of E. coli O157:H7 and S. Muenchen. Waxing reduced populations E. coli O157:H7 and S. Muenchen by up to 1.48 log10 cfu/apple. Compared to untreated apples, treatment of apples with water or waxes resulted in significant (P < or = 0.05) reductions in populations of E. coli O157:H7 and S. Muenchen during storage at 2 degrees C. Reductions on waxed apples stored at 21 degrees C were not as marked compared to reductions on waxed apples stored at 2 degrees C. With the exception of one wax, drying temperature did not significantly influence populations of yeasts and molds. Mold populations were less affected by wax applications than were yeasts, and were detected in higher numbers on apples treated with three of the five waxes compared to populations recovered from untreated control apples. None of the waxes evaluated can be relied upon to kill or remove E. coli O157:H7 and Salmonella on apples.

Colony Count, Microbial↗
PubMed2004

Tomato fruit cuticular waxes and their effects on transpiration barrier properties: functional characterization of a mutant deficient in a very-long-chain fatty acid beta-ketoacyl-CoA synthase.

Cuticular waxes play a pivotal role in limiting transpirational water loss across the plant surface. The correlation between the chemical composition of the cuticular waxes and their function as a transpiration barrier is still unclear. In the present study, intact tomato fruits (Lycopersicon esculentum) are used, due to their astomatous surface, as a novel integrative approach to investigate this composition- function relationship: wax amounts and compositions of tomato were manipulated before measuring unbiased cuticular transpiration. First, successive mechanical and extractive wax-removal steps allowed the selective modification of epi- and intracuticular wax layers. The epicuticular film consisted exclusively of very-long-chain aliphatics, while the intracuticular compartment contained large quantities of pentacyclic triterpenoids as well. Second, applying reverse genetic techniques, a loss-of-function mutation with a transposon insertion in a very-long-chain fatty acid elongase beta-ketoacyl-CoA synthase was isolated and characterized. Mutant leaf and fruit waxes were deficient in n-alkanes and aldehydes with chain lengths beyond C30, while shorter chains and branched hydrocarbons were not affected. The mutant fruit wax also showed a significant increase in intracuticular triterpenoids. Removal of the epicuticular wax layer, accounting for one-third of the total wax coverage on wild-type fruits, had only moderate effects on transpiration. By contrast, reduction of the intracuticular aliphatics in the mutant to approximately 50% caused a 4-fold increase in permeability. Hence, the main portion of the transpiration barrier is located in the intracuticular wax layer, largely determined by the aliphatic constituents, but modified by the presence of triterpenoids, whereas epicuticular aliphatics play a minor role.

3-Oxoacyl-(Acyl-Carrier-Protein) Synthase↗
PubMed1980

[Thermal and viscoelastic properties and problems on measurement methods of JIS for inlay waxes (author's transl)].

The thermal and viscoelastic properties of commercial inlay waxes were investigated by measurements of JIS (Japanese Industrial Standard), differential thermal analysis, dilatometry and dynamic viscoelasticity. The crystal transition from orthorhombic to hexagonal caused by paraffin was observed in inlay waxes. The thermal expansion of waxes was influenced by crystal transition temperature. The cooling shrinkage of internal wax pattern is smaller in wax having larger expansion from 25 degrees C to 37 degrees C. The viscoelastic properties of inlay waxes shifted to low temperature region with decreasing in crystal transition temperature. The various properties of inlay waxes greatly changed at temperature of peak for loss tangent. There were some problems on measurement methods of JIS and the reexamination of thermal expansion and flow was suggested. Commercial inlay waxes, respectively, have various different properties, but from these results the qualities of waxes can not be determined so it's necessary to choose a suitable wax according to various conditions.

Elasticity↗
PubMed1998

Epicuticular wax accumulation and fatty acid elongation activities are induced during leaf development of leeks

Epicuticular wax production was evaluated along the length of expanding leek (Allium porrum L.) leaves to gain insight into the regulation of wax production. Leaf segments from the bottom to the top were analyzed for (a) wax composition and load; (b) microsomal fatty acid elongase, plastidial fatty acid synthase, and acyl-acyl carrier protein (ACP) thioesterase activities; and (c) tissue and cellular morphological changes. The level of total wax, which was low at the bottom, increased 23-fold along the length of the leaf, whereas accumulation of the hentriacontan-16-one increased more than 1000-fold. The onset of wax accumulation was not linked to cell elongation but, rather, occurred several centimeters above the leaf base. Peak microsomal fatty acid elongation activity preceded the onset of wax accumulation, and the maximum fatty acid synthase activity was coincident with the onset. The C16:0- and C18:0-ACP-hydrolyzing activities changed relatively little along the leaf, whereas C18:1-ACP-hydrolyzing activity increased slightly prior to the peak elongase activity. Electron micrographic analyses revealed that wax crystal formation was asynchronous among cells in the initial stages of wax deposition, and morphological changes in the cuticle and cell wall preceded the appearance of wax crystals. These studies demonstrated that wax production and microsomal fatty acid elongation activities were induced within a defined and identifiable region of the expanding leek leaf and provide the foundation for future molecular studies.

Journal Article↗
PubMed2005

Cuticular wax deposition in growing barley (Hordeum vulgare) leaves commences in relation to the point of emergence of epidermal cells from the sheaths of older leaves.

In grasses, leaf cells divide and expand within the sheaths of older leaves, where the micro-environment differs from the open atmosphere. By the time epidermal cells are displaced into the atmosphere, they must have a functional cuticle to minimize uncontrolled water loss. In the present study, gas chromatography and scanning electron microscopy were used to follow cuticular wax deposition along the growing leaf three of barley (Hordeum vulgare L.). 1-Hexacosanol (C(26) alcohol) comprised more than 75% of extractable cuticular wax and was used as a marker for wax deposition. There was no detectable wax along the first 20 mm from the point of leaf insertion. Deposition started within the distal portion of the elongation zone (23-45 mm) and continued beyond the point of leaf emergence from the sheath of leaf two. The region where wax deposition commenced shifted towards more proximal (basal) positions when the point of leaf emergence was lowered by stripping back part of the sheath. When relative humidity in the shoot environment was elevated from 70% (standard growth conditions) to 92-96% for up to 4 days prior to analysis, wax deposition did not change significantly. The results show that cuticular waxes are deposited along the growing grass leaf independent of cell age or developmental stage. Instead, the reference point for wax deposition appears to be the point of emergence of cells into the atmosphere. The possibility of changes in relative humidity between enclosed and emerged leaf regions triggering wax deposition is discussed.

Hordeum↗