The composition of eggplant (Solanum melongena L.) leaf cuticular waxes was investigated. Chemical composition was determined as the first step in recognizing the importance to insect pest resistance of eggplant leaf surface chemistry. Waxes were extracted by dipping leaves into dichloromethane for 40 s. Relatively large samples were then fractionated by 'flash chromatography' on silica gel. Compounds were identified on the basis of their mass spectra from GC-MS analysis and retention data from GC-FID analysis, and quantified on the basis of peak areas from GC-FID analysis. Straight-chain alkanes and methyl-branched alkanes were the most abundant wax components. Minor quantities of esters of fatty acids with triterpene and aliphatic alcohols, free triterpene and aliphatic alcohols, sterols, and free fatty acids were also present on the eggplant leaf surface. This method is suitable for analysis of complex mixtures of plant cuticular lipids. It is also applicable to separation of samples for biological tests on insects.
A comparative study of potato leaf sesquiterpenes was carried out. GC, GC-MS, and NMR analyses were used to identify and quantify the sesquiterpenes in the leaf surfaces of 10 potato (Solanum tuberosum) varieties. Two sesquiterpene alcohols and 17 sesquiterpene hydrocarbons were identified and quantitatively determined. The distribution of the sesquiterpenes was found to be variety-specific. The sesquiterpene contents of the different potato varieties were subjected to cluster and principal component analyses. The eight potato varieties of the main chemotype cluster were dominated by beta-caryophyllene (9-148 ng/cm2), germacrene D (2-46 ng/cm2), germacrene D-4-ol (0.4-31 ng/cm2), beta-sesquiphellandrene (1-34 ng/cm2), and an unknown sesquiterpene alcohol III (0.2-37 ng/cm2). Chemometric classification distinguished two varieties, Mila and Vistula, from a major cluster. The Vistula variety was distinguished from the others by its high contents of beta-caryophyllene, alpha-humulene, germacrene D, and germacrene D-4-ol and the Mila variety by beta-elemene, trans-alpha-bergamotene, (Z)-beta-farnesene, (E)-beta-farnesene, trans-beta-bergamotene, beta-sesquiphellandrene, and unknown sesquiterpene alcohols I, II, III.
The sampling behavior of the Colorado potato beetle Leptinotarsa decemlineata Say (CPB) involves examination of the surface of potato leaves. It has been suggested that leaf surface compounds (volatiles and cuticular waxes) may be involved in host-plant recognition, acceptance or discrimination. Here we report on the effect of leaf surface extracts of six Polish commercial potato varieties on CPB feeding. We tested the influence of potato leaf surface extracts on CPB adult and larval feeding, then separated the extracts with HPLC, and finally tested the effect of the HPLC-separated fractions on CPB feeding. The bioassays were performed using potato leaf discs deprived of their original surface compounds. Applied to test discs at concentrations ten times higher than natural (10 leaf area equivalent), the extracts deterred CPB adults and larvae from feeding. HPLC-separated fractions composed of alkanes, sesquiterpene hydrocarbons, wax esters, benzoic acid esters, fatty acid methyl, ethyl, isopropyl and phenylethyl esters, aldehydes, ketones, methyl ketones, fatty acids, primary alcohols, b-amyrin and sterols did not affect adult CPB feeding. Similarly, alkanes, sesquiterpene hydrocarbons, wax esters, methyl ketones, sesquiterpene alcohols and secondary alcohols had no effect on larval CPB feeding. The sterol fraction (cholesterol, b-sitosterol and stigmasterol) acted as a phagostimulant to CPB larvae. We isolated a fraction demonstrating a phagodeterrent effect on CPB adults and larvae. The qualitative composition of the deterrent fraction was quite similar in all potato extracts, but there were quantitative differences between the varieties. Much further work is needed to identify the compounds that can produce the deterrent effect.
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