Surface Composition of Myrmecophilic Plants: Cuticular Wax and Glandular Trichomes on Leaves of <Emphasis Type="Italic">Macaranga tanarius</Emphasis>

Primary plant surfaces, covered with cuticles consisting of cutin and waxes, are important substrates for interaction with insects. The composition of leaf surfaces of the myrmecophilic plant Macaranga tanarius was studied. The prenylated flavanone nymphaeol-C was identified in surface extracts and was localized exclusively in glandular trichomes on the abaxial leaf side. The epidermal pavement cells surrounding these trichomes were covered with a smooth film of epicuticular wax from which few small wax crystals protruded. The epicuticular wax amounted to approximately 8 μg cm⁻², corresponding to 85% of the wax load on the adaxial as well as the abaxial leaf sides. The epicuticular wax mixtures from both leaf surfaces contained more than 70% primary alcohols, 14% fatty acids, 2% aldehydes, and traces of alkyl acetates, with chain lengths ranging from C₂₀ to C₃₈. In contrast, the intracuticular wax layer was largely dominated by triterpenoid alcohols α-amyrin, β-amyrin, and lupeol. Consequently, these characteristic compounds are not available for direct contact with insects on the plant surface.     

Influence of Some Fatty Acids on Oviposition by the Bruchid Beetle, Callosobruchus maculatus

The cowpea seed beetle, Callosobruchus maculatus, will lay its eggs on many potential hosts and inert surfaces. Oviposition on glass beads is stimulated by coating them with individual fatty acids. Nevertheless, female beetles reject mung seeds less frequently than beads treated with either an extract of mung seeds or, especially, an extract of mung seeds plus oleic acid. The addition of oleic acid to the extract resulted in a change in the sequence of oviposition behavior, notably an increase in a raised body position indicative of hosts of low acceptability. Fatty acids are present in the epicuticular waxes of legume seeds; wax extract of mung bean contains 32.4% fatty acid and 14 alkanes, whereas a wax extract of chickpea contains 5% fatty acid and 18 alkanes. Thus, chickpea may be a less acceptable host for oviposition than mung bean because of physical differences and/or because of chemical differences, including a reduced total level of fatty acid or the high proportion of oleic acid it is reported to contain. It is concluded that an appropriate mixture of fatty acids in the epicuticular waxes stimulates oviposition but that an elevated level of oleic acid in conjunction with others is deterrent.     

Gas-chromatographic analysis of the aliphatic fractions of brown coal wax

The individual composition of the fractions of wax isolated from brown coal from the Sergeevskoe deposit was studied using gas chromatography. It was found that the aliphatic constituents of wax are C₁₄–C₄₂ n-alkanes with high coefficients of oddness, C₁₄–C₃₀ saturated alcohols, C₁₄–C₃₆ higher fatty acids, and esters. The contribution of unsaturated compounds was found. The structure and formation conditions of coal are considered.     

Caterpillar Footprints as Host Location Kairomones for <Emphasis Type="Italic">Cotesia marginiventris</Emphasis>: Persistence and Chemical Nature

Herbivores walking over the epicuticular wax layer of a plant may leave tracks that disclose their presence to hunting predators or parasitoids. The braconid wasp Cotesia marginiventris is a solitary parasitoid of young noctuid caterpillars. It can locate potential hosts from a distance by orienting toward the scent of herbivore-damaged plants. Upon landing on the caterpillars’ food plant, the female parasitoid searches for further cues (kairomones) that confirm the presence of a suitable host. In a previous study, we showed that C. marginiventris recognizes the chemical footprints of absent Spodoptera frugiperda caterpillars on a leaf. Here, we report on the persistence and chemical nature of this host location kairomone. In a series of behavioral assays, we confirmed that caterpillars of S. frugiperda leave chemical tracks that elicit characteristic antennation behavior in C. marginiventris for up to 2 days. Both hexane extracts of caterpillar footprints and of the larvae’s ventral cuticle induced antennation and contained almost identical long-chain hydrocarbons, thus suggesting the prolegs and claspers as the kairomones’ main source. A series of linear C₂₁ to C₃₂ alkanes accounted for ca 90% of all identified compounds. Female wasps showed significant antennation responses on leaves treated with a reconstructed blend of these n-alkanes. However, wasp responses were relatively weak. Therefore, we presume that minor compounds, such as monomethyl-branched alkanes, which were also found, may contribute additionally to host recognition.     

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-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.     

Aldonamide-type gemini surfactants: Synthesis, structural analysis, and biological properties

A new group of nonionics, so-called sugar surfactants having a gemini structure, N,N′-bisalkyl-N,N′-bis[(3-gluconylamido)propyl]ethylenediamines [bis(C _n GA): C _n =n-C₈H₁₇, n-C₁₂H₂₅], N,N′-bisdodecyl-N,N′-bis[(3-glucoheptonylamido)propyl]ethylenediamine [bis(C₁₂GH)], and N,N′-bisalkyl-N,N′-bis[(3-lactobionylamido)propyl]ethylenediamines [bis(C _n LA): C _n =n-C₈H₁₇, n-C₁₂H₂₅], were prepared in a convenient four-step procedure from easily accessible reagents. Their structure and purity were confirmed by means of elemental analysis, electrospray ionization MS (ESI-MS), and NMR spectra—¹H, ¹³C, ¹H−¹³C COSY, and distortionless enhancement by polarization transfer. All tested surfactants were practically nontoxic to gram-negative bacteria and fungi, but they inhibited the growth of some gram-positive bacteria. From the results of the Closed Bottle test (OECD Guideline 301D) for biodegradability measurements, it was concluded that the tested aldonamide gemini structures are biodegraded by environmental microorganisms to 16–55% of the initial levels by day 28, the extent depending on both the aldonamide type and the alkyl chain length. Consequently, N,N′-bisalkyl-N,N′-bis[(3-aldonylamido)propyl]ethylenediamines are a surfactant class having low ecotoxicity and fulfilling requirements desired from an ecological standpoint.     

Selective Synthesis of Higher Linear α-olefins over Cobalt Fischer–Tropsch Catalyst

An efficient approach for more selective synthesis of higher linear α-olefins was achieved by utilizing suitable reaction media in FTS reaction. About 41.4% average α-olefins content in C₅–C₂₅ fractions was obtained at = 5 g-cat h mol⁻¹ in n-C₁₀ solvent, which is markedly higher than the value (2.5%) obtained in 85% N₂ + 15% n-C₆.     

Biological activity of synthetic hydrocarbon mixtures of cuticular components of the female housefly (Musca domestica L.)

Mixtures of (Z)-9-alkenes (C₂₁-C₃₃),n-alkanes (C₂₁-C₃₃), and mono- and dimethylalkanes (C₂₇, C₂₉), as components of the cuticular lipids washed from the female housefly (Musca domestica L.), were synthesized and assayed for their biological activity on male houseflies. The (Z)-9-alkenes and their hydrogenatedn-alkanes were synthesized from jojoba wax components by the appropriate chain elongation. The methylalkanes were prepared by Grignard coupling reaction of the appropriate alkyl halides, catalyzed by Li₂CuCl₄. Six- to 7-day-old virgin male houseflies exhibited the highest mating strike activity toward 6- to 7-day-old virgin females. The mating strike activity of the synthetic hydrocarbons was studied by exposing 6- to 7-day old virgin males to petrol-ether-rinsed 1 to 2-day-old dead females treated with these chemicals. (Z)-9-Tricosene was the most active hydrocarbon tested when it was applied in amounts of 10 g to a washed dead female. Amounts of 5 g of (Z)-9-tricosene did not cause sexual activity in the males. Mixture of (Z)-9-alkenes showed low activity when applied at 10-g amounts. However, when mixtures were made of 5 g of (Z)-9-tricosene with 5 fig of each of certain mixtures of (Z)-9-alkenes, the striking activity became as high as that of 10 g of (Z)-9-tricosene. This might be a synergism effect. The mixtures ofn-alkanes, as well as the mono- and dimethylalkanes, showed moderate activity. When (Z)-9-tricosene was added to these materials no increase in male activity was observed.     

Recent progress in the manufacture of alkanesulfonates: Photosulfochlorination of single-chain length (C12−C16) n-alkanes using SO2Cl2 at high conversion rates in pure phase and in the presence of solvent

Increasing concentration of sulfuryl chloride during the photosulfochlorination reaction under visible light shows that under these conditions n-alkanes react at high conversion rates instead of the conversion rate of 15% reported in the literature. This photosulfochlorination with sulfuryl chloride leads to better and more interesting results compared with those of photosulfochlorination by gas mixture. Indeed, nearly total conversion of n-alkanes, specifically n-heptane, n-dodecane, n-tetradecane, and n-hexadecane, occurred in pure phase, with a quantitative global yield, a sulfochlorination-to-chlorination molar ratio of about 1, and a relative reactivity of secondary to primary hydrogen of about 2.5, at a reaction temperature of 30°C and a reaction time of 120 min, using 2×10⁻² mol L⁻¹ of pyridine. Under these conditions, no polysulfochlorinated compounds are detected. These results are further improved using chlorobenzene as the solvent, instead of benzene. Indeed, the sulfochlorination of n-heptane at a conversion rate of 80% in the presence of chlorobenzene leads also to a quantitative reaction yield, a higher RSO₂Cl/RCl molar ratio, and, as expected, a high selectivity of secondary over primary hydrogen. Under these conditions, sulfochlorination of long-chain n-alkanes leads to the highest RSO₂Cl/RCl molar ratio for n-dodecane, n-tetradecane, and n-hexadecane belonging to detergent range, and the value of the molar ratio for these is between 1.45 and 1.7. The isomeric distribution of sulfochlorinated compounds obtained during sulfochlorination in the presence of solvent resembles that of secondary alkanesul-fonates produced by sulfoxidation reaction, whereas that obtained in pure phase has a similar conversion rate, is rich in primary isomer, and thus is different from that of classic radical reactions such as photochlorination or photosulfochlorination with gas mixture.     

Epicuticular Wax Chemicals in Zea mays Influence Oviposition in Ostrinia nubilalis

The chemical basis of oviposition elicitation in a generalist herbivore was determined by examination of oviposition responses in Ostrinia nubilalis to corn (Zea mays) chemicals in two-choice laboratory bioassays. A pentane extract of corn leaves stimulated oviposition and the activity persisted for three days, indicating that oviposition in O. nubilalis is elicited by low-volatility chemicals. Chemicals in the extract were fractionated by column chromatography on Florisil, using a sequence of solvents of increasing polarity. Bioassays of Florisil fractions indicated that the stimulants were eluted with nonpolar solvents. Positive bioassay results with an extract prepared by dipping corn leaves in pentane for 20 sec for extraction of leaf surface chemicals suggested that some of the active material was present in the leaf epicuticle. Gas chromatographic analyses and comparisons with retention times of standards suggested the presence of several n-alkanes in the dip extract. Five n-alkanes—hexacosane, heptacosane, octacosane, nonacosane, and tritriacontane—known to be present in the epicuticle of corn leaves were bioassayed, and all five elicited oviposition responses. These results suggest that oviposition elicitation in O. nubilalis is influenced by the presence of n-alkanes in the host plant epicuticle.     

Structure-activity relationship of unsaturated fatty acids as mosquito ovipositional repellents

Various straight-chain unsaturated fatty acids from C₁₄ to C₂₄ were evaluated for their ovipositional repellency against gravid females of the southern house mosquitoCulex quinquefasciatus Say, and the relationship between the structures of the fatty acids and their ovipositional repellency was determined. A double bond withZ configuration was prerequisite for an unsaturated fatty acid to be highly repellent;E isomers were less active or even inactive. No relationship was found between the repellency and the number of double bonds in the unsaturated fatty acids. In C₁₈ monounsaturated fatty acids, (Z)-9 acid was more active than (Z)-11 and (Z)-6 acids, indicating that a double bond at the 9 position rendered an acid highly repellent. Among (Z)-9-alkenoic acids of different chain lengths, the most repellent was C₁₈ acid which was also more active than (Z)-11-C₂₀, (Z)-13-C₂₂, and (Z)-15-C₂₄ acids. Oleic[(Z)-9-octadecenoic]acid, which met all these criteria, was the most ovipositionally repellent among the unsaturated fatty acids tested.Diptera: Culicidae.     

Egg dispersion in codling moth: Influence of egg extract and of its fatty acid constituents

Codling moth females (Cydia pomonella, Lepidoptera: Tortricidae) (CM) usually lay single eggs and have a tendency to disperse. In a first experiment we observed that single females exposed to 20 apples distribute their eggs regularly among apples, suggesting a dispersive oviposition behavior. In a dual-choice situation, isolated females avoided oviposition on areas of cardboard treated with a methylene dichloride egg extract at the dose of ca. 1.0 egg equivalent/cm². A strong avoidance was obtained in response to a 20-fold dose, which was accompanied by a significant reduction of total oviposition. Seven major compounds found by GC analyses in the methylene dichloride extract of 2 to 3-day-old eggs were saturated or unsaturated C₁₄-C₁₈ straight-chain fatty acids: myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. This was confirmed by GC-MS analysis of an extract made by ethyl ether. Smaller amounts of three methyl esters were also identified as methyl myristate, methyl palmitate, and methyl stearate. A blend of the seven fatty acids (FA) mimicked rather well the avoidance provoked by the extract, and this avoidance was confirmed by choice between treated and untreated fruits by single females. The treatment of apples with the 7FA mixture induced an aggregative distribution of the eggs among apples. We also found that the amounts of fatty acids harvested in the extract depend on the egg age. Amounts of fatty acids increased until eggs were 4 days old and then decreased before hatching. In this paper we discuss the possible role of simple molecules, such as fatty acids and their esters, as semiochemicals indicative of juvenile tissues.     

Characterization of skin-surface lipids from the monkey (Macaca fascicularis)

Skin-surface lipids from the monkeyMacaca fascicularis are composed of sterol esters (38%), cholesterol (4%) and two types of wax diesters, identified as Type II (IIa and IIb, 17% and 40%, respectively). Type IIa contained diesters of 1,2-alkanediols esterified with two molecules of long-chain (C₁₄−C₃₄) fatty acids having straight and branched chains. In the diesters IIa, fatty acids shorter than C₁₉ predominated in position 1, and fatty acids longer than C₂₀ predominated in position 2. Type IIb contained diesters of 1,2-alkanediols esterified with C₄ and C₅ branched-chain fatty acids (predominantly isovaleric acid) at position 1 and long-chain (C₁₄−C₂₇) acids, having straight and branched chains, at position 2. The shortchain acids were converted to 2-nitrophenylhydrazides and analyzed by high-performance liquid chromatography (HPLC). Ammonia chemical ionization (CI)-gas chromatography (GC)-mass spectrometry (MS) resolved the intact diesters IIb into 12 peaks corresponding to molecular weights ranging from 597 to 748, and showed that the molecular species, such as C₂₁−C₁₆−C₅ (diol, fatty acid in position 2, fatty acid in position 1), C₂₂−C₁₆−C₅ and C₂₃−C₁₆−C₅, were prevalent. The fatty acids from both diesters were mostly (>98%) saturated. The 1,2-alkanediols from both diesters consisted of C₁₆−C₂₆ saturated straight- and branched-chain components. The acyl groups of sterol esters contained 86% C₁₄−C₃₄ branched-chain acids. The unsaturated fatty acids (5.4%) belonged to a straight-chain monoenoic series having extremely long chains (C₁₈−C₃₄). The branched-chain structures in the fatty acids and diols were iso and anteiso. These results show the species-specific profile for the skin-surface lipid synthesis.     

Cuticular lipid constituents of cabbage seedpod weevils and host plant oviposition sites as potential pheromones

The cuticular lipids of cabbage seedpod weevils (Ceutorrhynchus assimilis Payk.) and those at their oviposition site, i.e., the seed pods of the host plant (Brassica napus L.), were analyzed by chromatographic techniques in conjunction with mass spectrometry (MS). Long chain hydrocarbons were most abundant in both lipids; however, the seed pods ofB. napus containedn-alkanes only, whereasC. assimilis showed a predominance of dimethylalkanes over internally branched methylalkanes as well as iso- andn-alkanes. The amounts of ketones, secondary alcohols and aldehydes, the usual plant components that are unique in insects, occurred in cuticular lipids of both organisms in approximately the same ratio. The composition ofn-alkanes,n-ketones, secondaryn-alcohols, iso- and anteisoaldehydes and esterified primary anteiso-alcohols of wax esters was similar betweenC. assimilis andB. napus. In both sources, qualitative but not quantitative similarities were observed in the composition ofn-aldehydes and esterified primaryn- and iso-alcohols, respectively. The esterified fatty acids of wax esters fromB. napus were composed of roughly equal proportions of saturated branched and unbranched components. The esterified fatty acids of wax esters and steryl esters fromC. assimilis consisted of major proportions of saturated as well as unsaturatedn-compounds, whereas iso- and anteiso structures were present in minor proportions only. Free fatty acids and traces of ethyl esters of fatty acids found inC. assimilis were mainly composed of unsaturatedn-compounds.     

Proportions of C18∶1n−7 and C18∶1n−9 fatty acids in canola seedcoat surface and internal lipids

Lipids of canola seedcoats (Brassica napus L. andB. rapa L.) were prepared by surface washing and by complete extraction of seed coats with toluene. The major fatty acyl-containing triacylglycerols, wax esters and free fatty acids were separated by thin-layer chromatography prior to transesterification and analysis by gas-liquid chromatography. The proportion of C18∶1n−7 to C18∶1n−9 was higher in the extracted lipids than in the surface-washed lipids for all three classes.     

Nature and role of sexual pheromones emitted by males ofAcrolepiopsis assectella (LEP.)

Seven compounds that do not exist in the extracts from legs of males have been isolated in the hair-pencil extracts of maleAcrolepiopsis assectella. By combining techniques of GC-MS and GC-FT-IR, six of these compounds have been identified. They are sixn-alkanes: hexadecane (C₁₆), heptadecane (C₁₇), octadecane (C₁₈), nonadecane (C₁₉), eicosane (C₂₀), and heneicosane (C₂₁). Twelven-alkanes of the homologous series, from the C₁₄–C₂₅ compounds were presented to virgin females, mated females, and males. At the end of the scotophase, four of then-alkanes (C₁₆, C₁₇, C₁₉, C₂₁) present in the hair-pencil extract induced the virgin females to adopt the acceptance posture after having induced the virgin females to remain stationary. The two othern-alkanes (C₁₈ and C₂₀) present in the extract have less effect on the females similar to then-alkanes not present in the males. The blends tested do not seem to indicate any synergy between the most active compounds. The threen-alkanes with an odd number of carbons and the C₁₆ compound would thus be the principle components of the male pheromone ofA. assectella. As well as their role of female aphrodisiac, they tend to make males and fertilized females flee.     

Thermodynamic and structural analyses of the solid phases in multi-alkane mixtures similar to petroleum cuts at ambient temperature

Structural analyses were carried out by X-ray diffraction, at ambient temperature, on multi-alkane samples whose mole fraction distribution shows a shape of the ‘exponential decreasing’ type, as observed in petroleum cuts. Nine samples, whose normal-alkane number varies from 15 to 23, have been studied with mole fraction continuous distributions of normal-alkanes going from C₂₂-C₃₆ to C₁₄-C₃₆: each mixture differs from the previous sample by the addition of a lighter n-alkane. At the solid state, the multi-alkane solid samples C₂₂-C₃₆ and C₂₁-C₃₆ are two-phase, C₂₀-C₃₆ to C₁₅-C₃₆, three-phase, and the broader distribution C₁₄-C₃₆, four-phase. In these polyphase solid systems, whose heaviest n-alkane is always C₃₆, the average composition of the heavy and middle phases are constant and their structure are isostructural to the β′ ordered intermediate solid solution, observed in n-alkane binary or ternary molecular alloys; the mean carbon atom number of the light phase decreases as the global average carbon atom number of the synthetic mixtures in relation to the addition of light n-alkanes and its structure simultaneously evolves from the β′ ordered intermediate solid solution towards the β-RI(Fmmm) and the disordered solid solutions, observed in pure n-alkanes: the light n-alkane added between each distribution intercalates itself into the structure whose molecule stacking period (thickness) is compatible with its own carbon chain length, in order to reduce the molecular gaps.     

Identification of 1-Octacosanal and 6-Methoxy-2-Benzoxazolinone from Wheat as Ovipositional Stimulants for Hessian Fly, Mayetiola destructor

Bioassay-guided fractionations of a dichloromethane extract of wheat epicuticular wax allowed the identification of 1-octacosanal and 6-methoxy-2-benzoxazolinone (MBOA) as the major components that stimulate oviposition by the Hessian fly, Mayetiola destructor. These compounds were identified by their mass spectral fragmentation patterns and by comparison of their gas chromatographic retention times with synthetic samples. Synthetic samples of 1-octacosanal or MBOA stimulated significant oviposition when compared with solvent controls. In combination, these compounds elicited a synergistic effect on the number of eggs laid by females compared to when they were presented alone. In a choice bioassay, a mixture of synthetic 1-octacosanal and MBOA in the approximate concentrations determined to be present in one plant equivalent of crude extract stimulated the same amount of oviposition as one plant equivalent of extract. This showed that together these two compounds appear to be responsible for the major proportion of the ovipositional stimulatory activity of the wheat epicuticular wax extract. Comparison of the activity of five straight-chain primary aldehydes with chain lengths from C₂₂ to C₃₀ revealed a relationship between chain length and the number of eggs laid by female Hessian flies, with 1-hexacosanal and 1-heptacosanal the most active of the aldehydes tested.     

Leaf wax alcohols inCoincya (Brassicaceae)

Free and ester alcohol compositions have been determined for leaf waxes in ten taxa belonging to the genusCoincya (Brassicaceae) on the Iberian Peninsula (Spain and Portugal). Size of leaf wax alcohols in the genusCoincya varies between 20 and 31 carbon atoms. This series is dominated by alcohols with an even number of carbon atoms. The most abundant alcohols are C₂₄, C₂₆ and C₂₈ among even and C₂₅, C₂₇ and C₂₉ among odd alcohols, both in free alcohols and in wax esters.     

Chemical composition of the wax secreted by a scale insect (Ceroplastes pseudoceriferus Green)

The wax material in the secretion of a scale insect,Ceroplastes pseudoceriferus Green was analyzed chemically with special interest to the composition of higher fatty acids and higher alcohols. The wax consists of 34.2% fatty acids, 27.1% unsaponifiable matter and 29.5% resin acids. The fatty acids were found to be a complex mixture of 15 normal acids ranging from C₈ to C₃₂. Of these, octacosanoic, triacontanoic and dotriacontanoic acids comprise over 30% of the wax. Presence of relatively large amount of unsaturated fatty acids of the C₁₈ series (2.8% of the wax) is of particular interest. From the unsaponifiable fraction, only one saturated straight chain aleohol, bexacosanol, was detected (2.7% of the original wax). The other unsaponifiable matter was considered to be cyclic or branched carbon chain, and consisted of at least 12 to 20 compounds. The resin acid fraction was also found to be a complex mixture of at least 13 to 14 components.