Synthesis of oleyl oleate wax esters in Arabidopsis thaliana and Camelina sativa seed oil

Seed oil composed of wax esters with long‐chain monoenoic acyl moieties represents a high‐value commodity for industry. Such plant‐derived sperm oil‐like liquid wax esters are biodegradable and can have excellent properties for lubrication. In addition, wax ester oil may represent a superior substrate for biodiesel production. In this study, we demonstrate that the low‐input oil seed crop Camelina sativa can serve as a biotechnological platform for environmentally benign wax ester production. Two biosynthetic steps catalysed by a fatty alcohol‐forming acyl‐CoA reductase (FAR) and a wax ester synthase (WS) are sufficient to achieve wax ester accumulation from acyl‐CoA substrates. To produce plant‐derived sperm oil‐like liquid wax esters, the WS from Mus musculus (MmWS) or Simmondsia chinensis (ScWS) were expressed in combination with the FAR from Mus musculus (MmFAR1) or Marinobacter aquaeolei (MaFAR) in seeds of Arabidopsis thaliana and Camelina sativa. The three analysed enzyme combinations Oleo3:mCherry:MmFAR1?c/Oleo3:EYFP:MmWS, Oleo3:mCherry:MmFAR1?c/ScWS and MaFAR/ScWS showed differences in the wax ester molecular species profiles and overall biosynthetic performance. By expressing MaFAR/ScWS in Arabidopsis or Camelina up to 59% or 21% of the seed oil TAGs were replaced by wax esters, respectively. This combination also yielded wax ester molecular species with highest content of monounsaturated acyl moieties. Expression of the enzyme combinations in the Arabidopsis fae1 fad2 mutant background high in oleic acid resulted in wax ester accumulation enriched in oleyl oleate (18:1/18:1 > 60%), suggesting that similar values may be obtained with a Camelina high oleic acid line.     

Chemotaxonomic Approach to the Central Balkan Sedum Species Based on Distribution of Triterpenoids in Their Epicuticular Waxes

Triterpenoid distribution in epicuticular waxes of 20 central Balkan Sedum L. species and four out‐groups of genera: Hylotelephium H. Ohba , Crassula L., Echeveria DC., and Kalanchoe Adans . were investigated for chemotaxonomic purposes. Identification and quantification of wax triterpenoids were performed by GC/MS and GC‐FID analyses. Distribution of identified triterpenoids (oleanane, lupane, and taraxerane series), as a pattern in statistical analysis, indicated very good agreement with phylogeny and systematics, except members of series Rupestria Berger , in which case the distribution of triterpenoids did not support known classification in a satisfactory manner. According to the obtained clustering, Kalanchoe is shown as similar to Sedum samples, while the other out‐groups were clearly separated.     

Effect of acidic fog on needle surface and water relations of Picea abies

Young spruce trees [ Picea abies (L.) Karst.) exposed to acidic fog (pH 3) showed a disintegration of the epicuticular waxes of the current year's needles as compared with trees treated with a fog of pH 5. The fog treatment was followed by a period in which the trees received different water supply. Under water stress conditions, trees that had been exposed to the acidic fog showed significantly higher transpiration rates than control trees that were treated with a fog of pH 5. The water loss of the controls was 0 at the middle of the photoperiod, while the trees pretreated with acidic fog still showed a substantial transpiration rate at this time. Water loss of excised twigs measured in the dry atmosphere of a desiccator also provided evidence that the water holding capacity of needles pretreated with acidic fog was affected. There is evidence that particularly the cuticular transpiration was increased by pretreatment with acidic fog. The xylem water potential of twigs pretreated with acidic fog was significantly lower than that of twigs pretreated with a fog of pH 5, while the osmotic potential was not affected by the different fog treatments. It is suggested that in draught years trees with a damaged cuticle caused by acidic fog will be affected in their resistance against water stress. In this way acidic fog could be involved in forest decline now widely spread in Central Europe.     

Epicuticular wax alkanes ofEricaceae andEmpetrum from alpine and sub-alpine heaths in Austria

Alkane distribution patterns were determined in the epicuticular wax of the leaves of 13 species and a hybrid fromEricaceae and one species ofEmpetrum (Empetraceae). As chemotaxonomic indicators, the results are of limited use only. The most uniform genus wasRhododendron, the most heterogeneousVaccinium. The dominant effect of genetic over environmental factors was apparent in most cases.     

Structure of the cuticle of the common house cricket with reference to the location of lipids

Cuticle segments from the thorax, abdomen, and jumping legs of the house cricket. Acheta domesticus, were examined using histological techniques for light microscopy, scanning and transmission electron microscopy, and direct examination of frozen-fractured cuticle. The surface of untreated cuticle is covered by a lipid film which obscures fine surface detail. Standard EM preparative procedures, as well as washing the cuticle with ethanol before examination, remove this film exposing previously covered openings to dermal gland ducts and wax canals. An epicuticle, exocuticle, mesocuticle, endocuticle, and a deposition layer were present in all transverse sections of cuticle. Light microscopy showed that the exocuticle and mesocuticle are heavily impregnated with lipids, whereas there is little lipid associated with the endocuticle. Frozen-fractured cuticle clearly shows the ‘plywood’ structure of the meso- and endocuticle, while the exocuticle fractures as if it were a solid sheet. The epicuticle is composed of a dense homogeneous layer, cuticulin, outer epicuticle, and the outer membrane. Superficial wax was detected only in cuticle samples prepared using vinylcyclohexane dioxide as a polar dehydrant. The results were used to construct a comprehensive model of the cuticle of A. domesticus.     

Epicuticular waxes of Secale cereale and Triticale hexaploide leaves

Wax on leaves of rye and of hexaploid Triticale (60–70-day-old plants) contains hydrocarbons (6–8%), esters (10%), free alcohols (14-8%), free acids (3%), hentriacontane-14,16-dione (39–45%), 25 (S)-hydroxyhentriacontane-14,16-dione (13–11%) and unidentified (14–15%). Diesters (1–3%) are also present in rye wax. Compositions of hydrocarbons (C₂₇-C₃₃) and esters (C₂₈,C₅₈) are similar for both waxes. Free and combined alcohols of rye wax are mainly hexacosanol but alcohols of Triticale wax are mainly octacosanol. The composition of Triticale wax is close to that of its wheat parent Triticum durum (cv. Stewart 63). Esters of wax from ripe rye contain 58% of trans 2,3-unsaturated esters. *NRCC No. 14033.     

Isolation of n-paraffins and sec-alcohols from Isotachis japonica

Two homologous series of n-paraffins and sec-alcohols from C₂₀ to C₃₄ or C₃₅ were isolated a wax constituents of a liverwort, Isotachis japonica. In both series, odd-members were predominant compared with even-members: the ratio of the total amount of odd- to total amounts of even-members was 3·9 in the n-paraffin homologues, and 16·5 in the sec-alcohol homologues.     

Composition and structure of secondary fatty alcohols and ketones,isolated from Pyrus malus skin wax

A homologous series of long-chain secondary fatty alcohols from C₂₁ to C₂₉ (C₂₉ being Predominant) has been isolated from the skin wax of a Bulgarian apple variety, Bouhavitsa. The alcohols were found only in a free state. Long-chain fatty ketones, with C₂₉ again markedly prevalent, have been isolated for the first time from the skin wax of this, and another variety, Tetovka. The C₂₉-ketone from both samples was identified by means of MS as nonacosan-10-one. A complete similarity has been found in the relative amounts of C₂₉ in the mixtures of secondary alcohols, ketones and paraffins.     

Survival and behavior of Plutella xylostella larvae on cabbages with leaf waxes altered by treatment with S-ethyl dipropylthiocarbamate

Cabbages (Brassica oleracea L.) treated with S-ethyl dipropylthiocarbamate (EPTC) herbicide had reduced amounts of leaf surface waxes (40.6% of controls) and reduced densities of leaf surface wax crystallites (20.8% of controls). Leaf waxes of EPTC-treated plants chemically and morphologically resembled leaf waxes of genetically glossy cabbages resistant to the diamondback moth Plutella xylostella (L.) (Lepidoptera: Plutellidae). Survival of larvae was significantly reduced on EPTC-treated cabbage plants in three out of four experiments (62.0–15.3% of survival on controls). P. xylostella neonates also moved more rapidly on EPTC-treated plants than on untreated controls (1.84±0.16 cm/min on controls vs. 3.94±0.24 cm/min on treated plants; P=0.0001). These results support the hypotheses that reduction in leaf waxes is the basis of resistance to P. xylostella in genetically glossy plants and that reduced acceptance by larvae is associated with this resistance. Modification of leaf surface waxes with EPTC or similar compounds may have potential as an economic control for P. xylostella in Brassica crops.