Ichtyotoxic activity of extracts from Mexican marine macroalgae

Seventy-three species of macroalgae from the Mexican Pacific, Atlantic and Caribbean coast were screened for ichtyotoxic activity. Ethanolic, acetonic and aqueous extracts were prepared and tested against the fish Carassius auratus. The extracts were classified on the basis of their effects as: toxic if the fish died in two hours or less; moderately toxic, if the organism behaved abnormally but death did notoccur, and non-toxic if the fish did not display any change. 79% species were ichtyotoxic to some degree. Extracts of 39 species were toxic, with at least one extract with lethal effects, 19 were moderately toxic and 15 species were non-toxic. Only the extracts ofDictyota bartayresiana, Dictyota cervicornis,Lobophora variegata, Bryothamnion triquetrum and Laurencia obtusa were toxic in all three solvents. The acetone and ethanol extracts were more active, and therefore are more suitable for extraction of toxic substances. This revised version was published online in August 2006 with corrections to the Cover Date.     

Biological activities of extracts from traditionally used Nigerian plants against the European corn borer, Ostrinia nubilalis

Preliminary investigations with ethanolic (EtOH) extracts from five Nigerian plants show that extracts of Piper guineense Schum and Thonn (Piperaceae), Cedrela odorata L. (Meliaceae), Dennettia tripetala G. Baker (Annonaceae) and Aframomum melegueta (Rosch) K. Schum (Zingiberaceae) in artificial diets significantly reduced larval growth of European corn borer (ECB), Ostrinia nubilalis Hubner, at a concentration of 1000 ppm (0.1%). An extract of Xylopia aethiopica (Dunal) A. Rich (Annonaceae) was ineffective. When the extracts were subsequently incorporated into artificial diets at 300 ppm and offered to neaonates, larval mortality increased in the order A. melegueta (13%), D. tripetala (13%), P. guineense (27%), and C. odorata (48%). Larval and adult emergence periods increased with increasing concentration of P. guineense, C. odorata and D. tripetala indicating a toxic response. Nutritional indices for habituated third instar larvae with the two most promising plant extracts, P. guineense and C. odorata, showed that the efficiencies of conversion of digested food (ECD) was significantly reduced at 300 ppm suggesting a postdigestive toxicity of the extracts. P. guineense and C. odorata extracts show the best potential for development as botanical insecticides.     

Temporal and ontogenetic aspects of protein induction in foliage of the tomato, Lycopersicon esculentum

Damage to foliage of the tomato, Lycopersicon esculentum, causes the induction of proteinase inhibitors and of the oxidative enzymes polyphenol oxidase, peroxidase, and lipoxygenase. The time courses of induction of these proteins by feeding of two caterpillar species (Manduca sexta and Helicoverpa zea) were studied in a series of experiments. In another series of experiments, the effects of plant age on the inducibility of these proteins were studied. In the time course experiments, induction of proteinase inhibitors and oxidative enzymes in the damaged leaflet was rapid, with higher protein activities evident in damaged leaflets within 12–24 h of damage, depending on the enzyme and the species of insect used to damage the plant. Systemic induction of proteinase inhibitors was also rapid, but systemic induction of polyphenol oxidase was delayed relative to systemic induction of proteinase inhibitors, possibly because high constitutive polyphenol oxidase activities obscured expression of systemic induction at earlier time points. Lipoxygenase and peroxidase were not induced systemically. Induction of all proteins persisted for at least 21 days. In the phenology experiments, inducibility of all proteins decreased in magnitude and was less consistent as plants aged. The results of these experiments exemplify the numerous constraints on induction in tomato plants. Knowledge of these physiological constraints is important to an understanding of the ecological role and causal basis of induced resistance.     

Genistein as an endogenous natural substrate of acidic peroxidases in lupin hypocotyls

Crude steam distillate from Ocimum gratissimum sprayed onto infection courts on detached cocoa pods moments after inoculation with Phytophthora palmivora completely inhibited the pathogen and blackpod lesion development on 75% of the infection courts. Disease suppression obtained with the extract was comparable to that obtained with a 2% Kocide 101 suspension. In the field, the O. gratissimum extract also suppressed lesion development although to a significantly lower (P = 0.05) extent in comparison with Kocide 101. Blackpod lesion expansion rates of 3.80, 3.56, 2.71 and 0.78 cm/day, respectively, were associated with pods treated in the field with C. citratus extract, tap water, O. gratissimum extract and 2% Kocide 101. The extract from Cymbopogon citratus was also ineffective on detached pods. Sporangia of P. palmivora from sporu-lating blackpod lesions on both detached and non-detached pods lost their infectivity within 1 h of treatment with the O. gratissimum extract. This effect was superior to that obtained with Kocide 101. Fungitoxicity of the extract on pods, however, was lost within 3 h of application. Thus, despite its in vivo effectiveness as an eradicant, the O. gratissimum extract, in its present form, has limited utility as a protectant fungicide.     

Reaction intensification for biocatalytic production of polyphenolic natural product di-C-β-glucosides

Polyphenolic aglycones featuring two sugars individually attached via C-glycosidic linkage (di-C-glycosides) represent a rare class of plant natural products with unique physicochemical properties and biological activities. Natural scarcity of such di-C-glycosides limits their use-inspired exploration as pharmaceutical ingredients. Here, we show a biocatalytic process technology for reaction-intensified production of the di-C-β-glucosides of two representative phenol substrates, phloretin (a natural flavonoid) and phenyl-trihydroxyacetophenone (a phenolic synthon for synthesis), from sucrose. The synthesis proceeds via an iterative two-fold C-glycosylation of the respective aglycone, supplied as inclusion complex with 2-hydroxypropyl β-cyclodextrin for enhanced water solubility of up to 50 mmol/L, catalyzed by a kumquat di-C-glycosyltransferase (di-CGT), and it uses UDP-Glc provided in situ from sucrose by a soybean sucrose synthase, with catalytic amounts (≤3 mol%) of UDP added. Time course analysis reveals the second C-glycosylation as rate-limiting (0.4–0.5 mmol/L/min) for the di-C-glucoside production. With internal supply from sucrose keeping the UDP-Glc at a constant steady-state concentration (≥50% of the UDP added) during the reaction, the di-C-glycosylation is driven to completion (≥95% yield). Contrary to the mono-C-glucoside intermediate which is stable, the di-C-glucoside requires the addition of reducing agent (10 mmol/L 2-mercaptoethanol) to prevent its decomposition during the synthesis. Both di-C-glucosides are isolated from the reaction mixtures in excellent purity (≥95%), and their expected structures are confirmed by NMR. Collectively, this study demonstrates efficient glycosyltransferase cascade reaction for flexible use in natural product di-C-β-glucoside synthesis from expedient substrates.     

Isolation of anti-fungal phenolic compounds from petioles of two Hevea brasiliensis (rubber) genotypes and their effect on Phytophthora meadii

Phenolic compounds were present in greater amounts in non‐infected petioles of genotypes of Hevea brasiliensis that are resistant to Phytophthora leaf disease than in genotypes that are susceptible. Phenolic compounds extracted from petioles of either susceptible (PB86) or resistant (RRIC100) genotypes, before or after infection with Phytophthora meadii, had anti‐fungal properties. Artificially infected petioles of PB86 had phenolic acids, triterpenoids or flavonoids, whereas healthy petioles contained only triterpenoids or flavonoids. However, healthy or infected petioles of RRIC100 contained only trace amounts of the above compounds and of vanillin (3‐methoxy‐4‐hydroxybenzaldehyde). Vanillin and umbelliferone (7‐hydroxycoumarin) were shown to suppress zoospore germination of P. meadii on glass slides and to inhibit its growth in pea broth and V‐8 juice agar. Vanillin was slightly more active than umbelliferone. Resistance of RRIC100 to Phytophthora was suspected as being related to the polymerisation of phenolic compounds to form lignin, which may suppress further spread of the pathogen's mycelium into healthy tissues. Formation of lignin from phenolic aldehydes as a barrier to disease spread may be a critical factor in resistance.     

The influence of macrophytes on a phytoplankton community in experimental conditions

The impact of submerged macrophytes or their extracts on planktonic algae was studied under experimental conditions. Live Ceratophyllum demersum L., its extract, and extracts of four other plant species induced modifications in the phytoplankton dominance structure. These modifications were: a decline in the number of Oscillatoria limnetica Lemm., which was the most numerous cyanobacterian species, and a decline in biomass and percentage contribution of all cyanobacteria to total algal biomass. This was accompanied by an increase in biomass and percentage contribution of green algae, especially Chlorella sp. and Chlamydomonas sp. Also, there was an increase in biomass and percentage contribution of nanoplankton (under 50 µm) to total phytoplankton biomass.The isolation of planktonic algae from direct influence of C. demersum by means of dialysis membranes caused an increase in number, biomass and percentage contribution of cyanobacteria. Release of organic compounds of over 3000 daltons by macrophytes apparently contributed to a decline of cyanobacteria by changing the phytoplankton dominance structure.     

Properties of polyphenol oxidase from tubers of the yam Dioscorea bulbifera

The subunit MW of Dioscorea bulbifera polyphenol oxidase (MW 115 000 ± 2000) determined by SDS-PAGE is ca. 31 000 indicating that the enzyme is an oligomeric protein with four subunits. K_i values of various inhibitors and their modes of inhibition have been determined with catechol and pyrogallol as substrates. p-Nitrophenol, p-cresol, quinoline and resorcinol are competitive inhibitors of catechol binding while only orcinol and p-nitrophenol behave in the same way towards pyrogallol as substrate. From the effect of pH on V_max, groups with pK values ca. 4.7 and 6.8 have been identified to be involved in catalytic activity. The Arrhenius activation energy (E_a) at pH 4.0 is 8.9 kcal/mol between 40–65°. At pH 7.0, the value is 22.1 kcal/mol between 40 and 60°. The enthalpies (ΔH) at pH 4.0 and pH 7.0 are 2.3 kcal/mol and 32.4 kcal/mol respectively. The results are discussed considering the conformational changes of the enzyme during substrate binding.     

Host fruit chemical stimuli eliciting distinct ovipositional responses from sibling species of Rhagoletis fruit flies

Laboratory experiments tested whether two economically-important sibling species of tephritid fruit flies have evolved distinct egg-laying responses to chemical stimuli on the fruits of their respective hostplants. The egg-laying preferences displayed by apple maggot flies, R. pomonella, and blueberry maggot flies, R. mendax, on artificial fruits treated with apple and blueberry extract paralleled their egg-laying responses to whole apples and blueberries. R. pomonella flies laid more eggs than R. mendax flies in artificial fruits treated with extract from ripe McIntosh apples, and vice versa for artificial fruits treated with extract from ripe Bluehaven blueberries. Furthermore, both species laid more eggs in artificial fruits treated with extract from their respective host fruits than control artificial fruits which were not treated with fruit extract. Prior electroantennogram recordings from R. mendax and R. pomonella flies exposed to volatiles from pentane extracts of apples and blueberries indicate that the antennal sensitivity of both species is selectively tuned to their respective host fruit odors. This differentiation in their olfactory responses to fruit odors could be important in mediating their distinct ovipositional responses to blueberry and apple fruits. Extract from unripe McIntosh apples also elicited egg laying by R. pomonella flies, however, artificial fruits treated with unripe apple extract received 1.9 times fewer eggs than those treated with ripe apple extract. Moreover, the numbers of R. pomonella ovipositor punctures and eggs placed in wax artificial fruits were increased when the artificial fruits were treated with a blend of 7 identified apple esters. Black coloration on these artificial fruits and the presence of apple esters had a synergistic effect on the egg-laying behavior of R. pomonella flies, which caused them to lay substantially more eggs per black fruit than white fruit treated with the same concentration of apple esters. In summary, our results indicate that the egg-laying responses of R. pomonella flies are mediated by the integration of information from fruit chemical and visual cues, and that R. mendax and R. pomonella flies have evolved divergent egg-laying responses to chemical stimuli on the fruits of their respective hostplants. These findings are discussed in the context of other studies on plant compounds which influence the ovipositional behavior of phytophagous Diptera.

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Stimuli chimiques des pommes et des myrtilles induisant la ponte des espèces jumelles, Rhagoletis pomonella et R. mendax

Résumé Des fruits artificiels en cire traités avec des extraits de fruits ont provoqué chez les espèces jumelles de R. mendax (Curran) et R. pomonella (Walsh) des réactions de ponte différentes suivant les stimulations chimiques par les fruits. Le comportement de ponte sur des fruits artificiels traités avec des extraits au pentane des myrtilles mûres (Vaccinium corymbosum L.) et de pommes mûres (Malus pumila Miller = Pyrus malus L.), est le même que sur des fruits naturels, ce qui montre que la réponse aux stimulations chimiques provenant du fruit constitue un aspect important de la reconnaissance de l'hôte. R. pomonella pond plus d'ufs que R. mendax sur les fruits artificiels traités à l'extrait de pommes mûres; c'est l'inverse pour les fruits traités aux extraits de myrtille. Les fruits artificiels traités avec des pommes ou des myrtilles provoquent la ponte de R. pomonella, tandis que les myrtilles mûres seules provoquent la ponte de R. mendax. Les extraits de pommes vertes stimulent la ponte de R. pomonella mais elle est alors 2 fois plus faible qu'avec des extraits de pommes mûres. Un mélange de 7 esters identifiés dans l'extrait de pomme induit aussi la ponte de R. pomonella. Le nombre de piqûres de tarièresfli dans les fruits artificiels en cire et le nombre d'ufs par fruit ont été augmentés par addition d'esters de pommes à des fruits blancs ou noirs. La couleur des fruits artificiels influence aussi la réaction de ponte de R. pomonella; la fréquence des piqûres de tarière contenant un uf et le nombre d'ufs par fruit étaient significativement plus élevés sur les fruits noirs que sur les fruits blancs traités avec la même concentration d'esters de pomme. Les fruits artificiels noirs traités avec la concentration la plus stimulante d'esters de pommes ont reçu 2, 3 fois plus d'ufs que les fruits blancs avec les mêmes concentrations en esters. Ces résultats montrent que les esters de pomme et la couleur noire stimulent synergiquement la ponte de R. pomonella sur des fruits artificiels.

     

The role of male accessory-gland substance on female reproduction with some observations of spermatogenesis in the stable fly

The bulbous tubular portion of the median ejaculatory duct functions as the accessory gland in the male stable fly, Stomoxys calcitrans. The gland started to enlarge after emergence when the fly was fed on sugar-water or blood. Implants of accessory glands from sugar-water or blood-fed males were effective in stimulating oviposition in virgin females. The injection into females of accessory-gland extracts from males fed blood prevented insemination; this extract was effective in concentration as low as 0.25 gland per female. Accessory-gland extracts from sugar-water fed males were only partially effective in preventing female insemination. However, the accessory-gland extracts of male stable flies had little effect on insemination when injected into three other species of female dipterans.Spermatogenesis was completed by the time of emergence, or shortly thereafter, a process independent of blood feeding. Three types of spermatids were identified, forming a continuum of spermiogenesis stages. Fat, fusiform spermatozoa elongated to become thin, elongate and mature spermatozoa.