Phytochemical and biological investigation of <Emphasis Type="Italic">Stevia rebaudiana</Emphasis> Bertoni; 1-labdane-type diterpene

Five labdane diterpenoids, austroinulin, iso-austroinulin, sterebin E, sterebin E acetate, and sterebin A acetate, along with hydrocarbons, aliphatic alcohols, β-amyrin, β-sitosterol and stigmasterol were isolated from the chloroform soluble fraction of the methanol extract of Stevia rebaudiana leaves. Chlorogenic and caffeic acids were isolated from the EtOAc fraction. All the isolated compounds were identified using spectral tools. The chloroform and methanol extracts proved significant anti-inflammatory effect and caused marked inhibition of carrageenan-induced paw oedema in rats.     

Allelochemical resistance of bald cypress,Taxodium distichum,heartwood to the subterranean termite,Coptotermes formosanus

The heartwood of bald cypress,Taxodium distichum (L.) Rich., resisted feeding attack by the Formosan subterranean termite,Coptotermes formosanus Shiraki. Hexane-extracted heartwood, however, was consumed at > 12 times the amount of sound heartwood eaten. A bioassay usingT. distichum sapwood as a feeding substrate was employed to assess the antitermitic activity of successive hexane, acetone, and methanol extracts of heartwood shavings and isolates derived from the active hexane extract. Two fractions, eluted from the crude hexane extract by liquid chromatography, significantly reduced termite feeding compared to the parent extract, while a third fraction was less active than the original hexane extract. Each fraction contained one major component. All three components were structurally related diterpenes. The two most active heartwood constituents were identified by GC-MS and NMR as ferruginol and manool, while the third and least active, but most prevalent, compound in heartwood was identified as nezukol. Results of bioassays suggest that these allelochemicals act principally as feeding deterrents with accompanying termite mortality due to starvation.     

Sphaeropsidin A: A Pimarane Diterpene with Interesting Biological Activities and Promising Practical Applications

Sphaeropsidin A (SphA) is a tetracyclic pimarane diterpene, first isolated as the main phytotoxin produced by Diplodia cupressi the causal agent of a severe canker disease of Italian cypress (Cupressus sempervirens L.). It was also produced, together with several analogues, by different pathogenic Diplodia species and other fungi and showed a broad array of biological activities suggesting its promising application in agriculture and medicine. The anticancer activity of SphA is very potent and cell specific. Recent studies have revealed its unique mode of action. This minireview reports the structures of SphA and its family of natural analogues, their biosynthetic origins, their fungal sources, and biological activities. The preparation of various SphA derivatives is also described as well as the results of structure-activity relationship (SAR) studies and on their potential practical applications.     

Pseudopterosin Content Variability of the Purple Sea Whip Pseudopterogorgia elisabethae at the Islands of San Andres and Providencia (Sw Caribbean)

To determine pseudopterosin composition and concentration in colonies of Pseudopterogorgia elisabethae from the islands of San Andres and Providencia, we collected fragments of individual colonies at various sites and depth ranges around the islands. Chromatographic profiles of the polar fraction, particularly those obtained by HPLC-MS analyses, allowed us to recognize two different chemotypes. Chemotype 1 characterized samples from Providencia whereas chemotype 2 characterized samples from San Andres. A complex pseudopterosin mixture (compounds 1-13) characterized chemotype 1. These compounds were isolated by a combination of chromatographic methods and identified by spectroscopic methods (MS, UV, 1H, and 13C NMR). We identified the known pseudopterosins G and K and seco-pseudopterosin A. We also isolated and identified seven new compounds, pseudopterosins P-V, isomers of known pseudopterosins. Pseudopterosins G and K were found at concentrations ranging between 1 and 3% of the animal dry mass. Pseudopterosins Q and U were the major compounds reaching up to 6% of the animal dry mass at some locations. Major metabolites in chemotype 2 had a molecular weight and fragmentation pattern different from that observed in the pseudopterosins, as determined by HPLC-MS. Total pseudopterosin concentration in this chemotype was below 3% dry mass at all sites. Total pseudopterosin concentration was significantly higher in chemotype 1, with concentrations ranging between 4 and 20% dry mass. At most locations on Providencia, however, total pseudopterosin concentration ranged between 11 and 15% dry mass. Concentrations exceed reports from other locations in the Caribbean. Furthermore, pseudopterosin composition in our samples is quite different from those in specimens of P. elisabethae from the Bahamas and Bermuda. Pseudopterosins G, K, and P-V are characteristic of P. elisabethae colonies from the island of Providencia, while pseudopterosins A-D are characteristic of colonies of P. elisabethae from the Bahamas islands, and pseudopterosins E-L have been isolated from P. elisabethae from the Bahamas and Bermuda. The overall morphology of P. elisabethae can be variable, and chemical differences are not correlated to specific morphs. We confirmed the species identity of each colony by morphological and sclerite analysis and found no significant differences in sclerite dimensions among different colonies and chemotypes.     

Effects of cuticular duvane diterpenes from green tobacco leaves on tobacco budworm (Lepidoptera: Noctuidae) oviposition

Five cuticular chemical components isolated from the green leaves of tobacco introductions (TIs) and a commercial tobacco cultivar were tested for their effects on tobacco budworm,Heliothis virescens (F), oviposition in cage bioassays, and field experiments. These chemicals were sprayed onto budworm-resistant TI 1112 tobacco which produces low levels of most cuticular components. Individual duvane diterpenes (- and -4,8,13-duvatrien-1-ols and - and -4,8,13-duvatriene-1,3-diols) increased tobacco budworm egg laying on sprayed TI 1112 plants.cis-Abienol, docosanol, and docosanyl myristate were inactive.Mention of a company or proprietary product does not imply endorsement or recommendation by the U.S. Department of Agriculture. Paper No. 9702 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina 27695-7601.     

Interspecific Variation in Defense Secretions of Malaysian Termites from the Genus <Emphasis Type="Italic">Bulbitermes</Emphasis>

The chemicals of the defense secretions of Malaysian Bulbitermes, B. singaporensis, B. germanus, B. sarawakensis, and Bulbitermes sp. B, show that B. singaporensis is distinct from the other species, which are themselves closely related; the genetic distance between B. singaporensis and B.germanus is 0.71. B. singaporensis contains tetracyclic kempane, and B. germanus and B. sarawakensis contain tricyclic trinervitene; Bulbitermes sp. B contains a mixture of kempane and trinervitene. The mono- and diterpenoid compositions are species-specific.     

Variability of terpene content in the soft coralSinularia flexibilis (Coelenterata: Octocorallia), and its ecological implications

Colonies of the soft coralSinularia flexibilis (Quoy & Gaimard) (Coelenterata, Octocorallia) were collected at Lizard Island (14°40S and 145°28E) Research Station. Extraction of the corals and quantitative chemical analysis for the three major diterpene components, flexibilide, dihydroflexibilide, and sinulariolide, afforded average ratios of 431 respectively. Colonies, sized on the basis of the sterile stalk circumference, were analyzed for possible correlations between size and chemical composition. The major metabolite, flexibilide, was inversely correlated with colony size, while sinulariolide concentration showed a direct correlation. The concentration of dihydroflexibilide was independent of colony size. Samples were further analyzed with respect to site of collection. Colonies were collected at three distinct reefal sites. One was characterized by large monospecific stands ofParites cylindrica, a second was a sandy bottom site with a mixed community of soft corals and occasional scleractinians, while the third site was a very diverse reef community with many species of scleractinian corals.Sinularia flexibilis was well represented at each site, and the concentration of flexibilide and sinulariolide varied significantly among sites. The concentration of flexibilide was significantly higher at the third, highly competitive site, while the concentration of sinulariolide was highest at thePorites-dominated site. Dihydroflexibilide levels were independent of site. It seems likely that concentrations of flexibilide, a highly cytotoxic molecule involved in interference competition, and sinulariolide, a known algicide probably responsible for colony maintenance, may be influenced by their environments.     

Correspondence of soldier defense secretion mixtures with cuticular hydrocarbon phenotypes for chemotaxonomy of the termite genus Reticulitermes in North America

Soldier defense secretions from samples of Reticulitermes collected in California, Nevada, Arizona, New Mexico, and Georgia were characterized and correlated with cuticular hydrocarbon phenotypes. Twenty-seven cuticular hydrocarbon phenotypes have been defined, and soldier defense secretion (SDS) phenotypes have been described for 25 of these. Forty-five terpenoid compounds were found, including monoterpenes, sesquiterpenes, and a few diterpenes. The monoterpenes include (–)--pinene, (–)--pinene, (–)-camphene, myrcene, (Z)- and (E)-ocimene, and (–)-limonene. The major sesquiterpenes produced are (+)--cadinene, (+)--cadinene aldehyde, (–)-germacrene A, germacrene B, -himachalene, and -bisabolene. Some SDS phenotypes pair with more than one cuticular hydrocarbon phenotype; however, with two exceptions, each hydrocarbon phenotype is associated with only one SDS phenotype. These chemical characterizations lend support to the conclusion that there are numerous undescribed species of Reticulitermes in North America.     

Leaf surface chemicals fromNicotiana affecting germination ofPeronospora tabacina (adam) sporangia

A bioassay was used to evaluate the effects of cuticular leaf components, isolated fromN. tabacum, N. glutinosa (accessions 24 and 24a), and 23other Nicotiana species, on germinationof P. tabacina (blue mold). The leaf surface compounds included- and-4,8,13,-duvatriene-l,3-diols (DVT-diols), (13-E)-labda-13-ene-8-,15-diol (labdenediol), (12-Z)-labda-12,14-diene-8-ol (cis-abienol), (13-R)-labda-8,14-diene-13-ol (manool), 2-hydroxymanool, a mixture of (13-R)-labda-14-ene-8,13-diol (sclareol), and (13-S)-labda-14-ene-8,13-diol (episclareol), and various glucose and/or sucrose ester isolates. The above in acetone were applied onto leaf disks of the blue moldsusceptibleN. tabacum cv. TI 1406, which was then inoculated with blue mold sporangia. Estimated IC₅₀ values (inhibitory concentration) were 3.0g/cm² for-DVT-diol, 2.9/cm² for-DVT-diol, 0.4g/cm² for labdenediol and 4.7g/cm² for the sclareol mixture. Manool, 2-hydroxymanool, andcis-abienol at application rates up to 30g/cm² had little or no effect on sporangium germination. Glucose and/or sucrose ester isolates from the cuticular leaf extracts of 23Nicotiana species and three different fractions fromN. bigelovii were also evaluated for antimicrobial activity at a concentration of 30g/cm². Germination was inhibited by >20% when exposed to sugar esters isolated fromN. acuminata, N. benthamiana, N. attenuata, N. clevelandii, andN. miersii, and accessions 10 and 12 ofN. bigelovii. These results imply that a number of compounds may influence resistance to blue mold in tobacco.     

The formation of phenolic diterpenes in Rosmarinus officinalis L. under Mediterranean climate

 The formation of phenolic diterpenes in the leaves of rosemary plants grown under Mediterranean climate was investigated. Low precipitation coincided with high solar radiation during the summer, resulting in water and light stress of the plants. The highest concentrations of the major diterpenes carnosic acid and carnosol were found during the winter and the lowest concentrations during the summer, which is likely to be the result of their enzymatic conversion in the presence of singlet oxygen. All other diterpenes followed different trends during the season. The concentration of carnosic acid increased with the relative water content of the leaf but decreased with increasing solar radiation and temperature. 11,12-di-O-Methylisorosmanol was found for the first time in rosemary leaves. Received: 22 March 1999