Location, location, location: Feeding site affects aphid performance by altering access and quality of nutrients.

Aphid feeding behavior and performance on a given host plant are influenced by the plants' physical and chemical traits, including structural characters such as trichomes and nutritional composition. In this study, we determined the feeding behavior and performance of soybean aphids (Aphis glycines) on the stem, the adaxial (upper), and the abaxial (lower) leaf surfaces during early vegetative growth of soybean plants. Using the electrical penetration graph technique, we found that aphids feeding on the stem took the longest time to begin probing. Once aphids began probing, the sieve elements were more conducive to feeding, as evidenced by less salivation on the stem than either leaf surface. In whole-plant assays, stems harbored higher aphid populations, and aphids had shorter development time on stems than the adaxial and the abaxial leaf surfaces. We compared trichome density and length on the stem, the adaxial, and the abaxial leaf surfaces to investigate whether plant trichomes affected aphid feeding and performance. There were higher density and longer trichomes on stems, which likely resulted in aphids taking a longer time to probe. Still a negative impact on aphid population growth was not observed. Analysis of phloem sap composition revealed that vascular sap-enriched exudates from stems had higher sugars and amino acids than exudates from leaves. In artificial diet feeding assays, the population of aphids reared on a diet supplemented with stem exudates was higher than on a diet supplemented with leaf petiole exudates which is in agreement with results of the whole-plant assays. In summary, our findings suggest that the performance of soybean aphids on a specific plant location is primarily driven by accessibility and the quality of phloem composition rather than structural traits.

Effects of Aqueous Extracts from Amazon Plants on Plutella xylostella (Lepidoptera: Plutellidae) and Brevicoryne brassicae (Homoptera: Aphididae) in Laboratory, Semifield, and field trials.

We evaluated the insecticide activities of aqueous extracts of five species of plants from the Ecuadorian Amazon (Deguelia utilis (ACSm.) AMGAZEVEDO (Leguminosae: Papilionoideae), Xanthosoma purpuratum K. Krause (Alismatales: Araceae), Clibadium sp. (Asteracea: Asterales), Witheringia solanacea L'Hér (Solanales: Solanaceae), and Dieffenbachia costata H. Karst. ex Schott (Alismatales: Araceae)) plus Cymbopogon citratus Stapf. (Poales: Poaceae) under laboratory, open-field conditions in Plutella xylostella L. (diamondback moth), and semifield conditions in Brevicoryne brassicae L. Tap water was used as a negative control, and synthetic insecticides were used as positive controls. In a laboratory bioassay, aqueous extracts of D. utilis resulted in P. xylostella larval mortality. In contrast to chlorpyrifos, all botanicals were oviposition deterrents. All extracts except Clibadium sp. decreased leaf consumption by P. xylostella larvae. In semifield experiments, D. utilis, Clibadium sp., D. costata, and X. purpuratum initially controlled the population of B. brassicae, but 7 d after application, all botanicals except the D. utilis lost their ability to control the pest. In field experiments on broccoli crops in both dry and rainy seasons, the extracts did not control the abundance of P. xylostella, where as a mixture of two insecticides (chlorpyrifos + lambda cyhalothrin) did. These results show some incongruences from laboratory to semifield and field conditions, indicating that more studies, including the identification of the chemicals responsible for the biological activity, its stability, and the effects of chemotypes on insecticidal activity, are needed to understand the potential of these plant species as botanical insecticides.

Encapsulation of Mentha pulegium Essential Oil in Yeast Cell Microcarriers: An Approach to Environmentally Friendly Pesticides.

A green approach for the encapsulation of Mentha pulegium essential oil in commercial baker's yeast and its evaluation as a pesticide against the insect pest Myzus persicae are presented. Upon treating aqueous yeast cell dispersion with the essential oil, the formation of essential-oil-loaded microparticles of about 9 µm is observed, with a loading capacity ranging from 29 to 36%, depending upon the encapsulation conditions. The thermal properties of the microparticles were characterized using differential scanning calorimetry and thermogravimetric analysis, confirming the protection of the essential oil from the cells. Encapsulation prolonged the insecticidal activity of the essential oil by 3 days.

Synthesis and insecticidal activities of novel solanidine derivatives.

BACKGROUND: Potato (Solanum tuberosum) is the fourth culture in the world and is widely used in the agri-food industries. They generate by-products in which α-chaconine and α-solanine, the two major solanidine-based glycoalkaloids of potato, are present. As secondary metabolites, they play an important role in the protection system of potato and are involved in plant protection against insects. To add value to these by-products, we described here new glycoalkaloids that could have phytosanitary properties. RESULTS: Solanidine, as a renewable source, was modified with an azido linker and coupled by copper-catalyzed alkyne azide cycloaddition to alkynyl derivatives of the monosaccharides found in the natural potato glycoalkakoids: D-glucose, D-galactose and L-rhamnose. The efficacy of our compounds was evaluated on the potato aphid Macrosiphum euphorbiae. The synthetic compounds have stronger aphicidal properties against nymphs than unmodified solanidine. They also showed strong aphicidal activities on adults and a negative impact on fecundity. CONCLUSION: Our synthetic neoglycoalkaloids affected Macrosiphum euphorbiae survival at the nymphal stage as well as at the adult stage. Furthermore, they induced a decrease in fecundity. Our results show that chemical modifications of by-products may afford new sustainable compounds for crop and plant protection. © 2018 Society of Chemical Industry.

Effect of petroleum-derived substances on life history traits of bird cherry-oat aphid (Rhopalosiphum padi L.) and on the growth and chemical composition of winter wheat.

The aim of the study was to determine the effects of various petroleum-derived substances (PDSs), namely petrol, diesel fuel, and spent engine oil, on life history traits of the bird cherry-oat aphid Rhopalosiphum padi L., and on the growth and chemical composition of its host plant-winter wheat Triticum aestivum L. Each substance was tested separately, using two concentrations (9 and 18 g kg-1). Plants were cultivated in both control and contaminated soils. In early October 2013, soil was contaminated and after 1 week, winter wheat seeds, 'Batuta' cultivar, were sown. In early June 2014, observations of the effect of petroleum-derived substances on traits of three successive generations of aphids were conducted. Aphids were inoculated separately on leaves using cylindrical cages hermetically closed on both sides. Contamination of aphid occurred through its host plant. Results showed that all of the applied petroleum-derived substances have a generally adverse effect on the developmental parameters in aphids, resulting in the decrease of its fecundity, shortening its average life span, and most often lowering of the population intrinsic growth rate. PDSs caused the limitation of growth in wheat plants; whereas, changes in nutrient contents and heavy metals depended on the part of the plant analysed, the substance applied, and on its dose. The negative relationships between the contents of both some macro-elements (Ca, K, P) and heavy metals (Mn, Cd, Cu, and Zn) and the developmental parameters of particular generations of R. padi were observed. The high susceptibility of R. padi to the presence of PDSs in the substrate for the host plant should be emphasised-the clear-cut changes in the life span and fecundity, with relatively small changes in the chemical composition of the plant, constitute an evident indication that the developmental parameters of aphids have the potential for the use as bio-indicator to evaluate the state of the environment contaminated by PDSs.

Lethal and Sublethal Effects of Mineral Oil on Potato Pests.

Mineral oil is a product used to reduce Potato Virus Y transmission in potato fields. However, there is little information available about other effects that oil may have on insect pests of potato. To better understand how mineral oil affects potato pests, we performed a series of experiments testing the effects of oil on mortality, behavior, and development of potato aphids, Macrosiphum euphorbiae (Thomas) (Hemiptera: Aphididae), green peach aphids, Myzus persicae (Sulzer) (Hemiptera: Aphididae), and Colorado potato beetles, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). All three species showed negative behavioral responses to oil-treated potato foliage. Oil treatment also increased aphid mortality. Colorado potato beetle mortality was not affected, but developing on oil-treated potato plants resulted in prolonged development and smaller adults. Additionally, oil acted synergistically with the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae); Colorado potato beetle larvae were killed more rapidly when sprayed with both products compared with when sprayed with B. bassiana alone. Based on these results, mineral oil has the potential for expanded use in potato IPM programs.

Host Plant-Herbivore-Predator Interactions in Chrysoperla carnea (Neuroptera: Chrysopidae) and Myzus persicae (Homoptera: Aphididae) on Four Plant Species Under Laboratory Conditions.

The common green lacewing, Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), is a well-known biocontrol agent. The current study examined host plant-herbivore-predator interactions with C. carnea and Myzus persicae on four host plants (peach, almond, pepper, and potato). The experiments were carried out at 25 ± 1°C and 65 ± 5% RH at a photoperiod of 16:8 (L:D) h). Duration of the preadult growth period, adult longevity, fecundity, and population growth parameters were analyzed based on the age-stage, two-sex life table theory. The shortest and longest preadult developmental times of the predator were observed on the peach and potato, respectively. The highest and lowest predation rate, oviposition period, and male and female longevity of predator were also observed on the peach and potato, respectively. The lowest intrinsic rate of increase (r) and finite rate of increase (λ) were observed on the potato (0.1087 and 1.11 d-1, respectively) and the highest on the peach (0.1460 and 1.15 d-1, respectively). The maximum and minimum mean generation times (T) were 41.84 and 35.59 d in the potato and peach, respectively. Overall, peach was found to be a more appropriate host than the other host plants for development and predation fitness of C. carnea. These findings reveal that information on tritrophic interactions and subsequent life table evaluation of natural enemies improves integrated pest management programs.

Treatment by glyphosate-based herbicide alters life history parameters of the rose-grain aphid Metopolophium dirhodum.

Glyphosate is the number one herbicide in the world. We investigated the sub-lethal effects of this herbicide on the aphid Metopolophium dirhodum (Walker), using an age-stage, two-sex life table approach. Three concentrations of the herbicide (low - 33.5, medium - 66.9 and high - 133.8 mmol dm(-3) of active ingredient) and distilled water as the control were used. The LC50 of the IPA salt of glyphosate on M. dirhodum was equivalent to 174.9 mmol dm(-3) of the active ingredient (CI95: 153.0, 199.0). The population parameters were significantly negatively affected by herbicide application, and this negative effect was progressive with the increasing concentration of the herbicide. A difference of two orders of magnitude existed in the predicted population development of M. dirhodum between the high concentration of the herbicide and the control. This is the first study that comprehensively documents such a negative effect on the population of an herbivorous insect.

MOLECULAR CLONING, EXPRESSION PATTERN OF MULTIDRUG RESISTANCE ASSOCIATED PROTEIN 1 (MRP1, ABCC1) GENE, AND THE SYNERGISTIC EFFECTS OF VERAPAMIL ON TOXICITY OF TWO INSECTICIDES IN THE BIRD CHERRY-OAT APHID.

The ATP-binding cassette (ABC) transporters are important transmembrane proteins encoded by a supergene family. The majority of ABC proteins are primary active transporters that bind and hydrolyze ATP to mediate the efflux of a diverse range of substrates across lipid membranes. In this study, we cloned and characterized a putative multidrug resistance associated protein 1 (MRP1) from Rhopalosiphum padi encoded by ABCC1. Structural analysis showed that this protein has structural features typical of the ABC transporter family. Phylogenetic analysis indicated that the amino acid sequence was highly similar that of the corresponding protein from Acyrthosiphon pisum. Real-time quantitative polymerase chain reaction (PCR) analysis showed that ABCC1 was expressed throughout all R. padi developmental stages, with the highest level of expression in the fourth larval instar. We also examined ABCC1 expression in four different tissue types and found that it was most highly expressed in the midgut. Exposing R. padi to imidacloprid and chlorpyrifos increased ABCC1 expression. Furthermore, ABCC1 expression was higher in the imidacloprid-resistant (IR) and chlorpyrifos-resistant (CR) strains than in an insecticide-susceptible strain (SS) of R. padi. Exposing R. padi to verapamil in combination with insecticides significantly increased the toxicity of the insecticides. The respective synergy factor of CR and IR R. padi strain was 1.33 and 1.26, which was lower than that (2.72 and 1.64, respectively) of the SS. Our results clarify the biological function of ABCC1 in R. padi, particularly its role in insecticide resistance, and suggest novel strategies for pest management that use ABC transporter inhibitors to increase the effectiveness of insecticides.

The ethylene response factor Pti5 contributes to potato aphid resistance in tomato independent of ethylene signalling.

Ethylene response factors (ERFs) comprise a large family of transcription factors that regulate numerous biological processes including growth, development, and response to environmental stresses. Here, we report that Pti5, an ERF in tomato [Solanum lycopersicum (Linnaeus)] was transcriptionally upregulated in response to the potato aphid Macrosiphum euphorbiae (Thomas), and contributed to plant defences that limited the population growth of this phloem-feeding insect. Virus-induced gene silencing of Pti5 enhanced aphid population growth on tomato, both on an aphid-susceptible cultivar and on a near-isogenic genotype that carried the Mi-1.2 resistance (R) gene. These results indicate that Pti5 contributes to basal resistance in susceptible plants and also can synergize with other R gene-mediated defences to limit aphid survival and reproduction. Although Pti5 contains the ERF motif, induction of this gene by aphids was independent of ethylene, since the ACC deaminase (ACD) transgene, which inhibits ethylene synthesis, did not diminish the responsiveness of Pti5 to aphid infestation. Furthermore, experiments with inhibitors of ethylene synthesis revealed that Pti5 and ethylene have distinctly different roles in plant responses to aphids. Whereas Pti5 contributed to antibiotic plant defences that limited aphid survival and reproduction on both resistant (Mi-1.2+) and susceptible (Mi-1.2-) genotypes, ethylene signalling promoted aphid infestation on susceptible plants but contributed to antixenotic defences that deterred the early stages of aphid host selection on resistant plants. These findings suggest that the antixenotic defences that inhibit aphid settling and the antibiotic defences that depress fecundity and promote mortality are regulated through different signalling pathways.

Complex effects of fertilization on plant and herbivore performance in the presence of a plant competitor and activated carbon.

Plant-herbivore interactions are influenced by host plant quality which in turn is affected by plant growth conditions. Competition is the major biotic and nutrient availability a major abiotic component of a plant's growth environment. Yet, surprisingly few studies have investigated impacts of competition and nutrient availability on herbivore performance and reciprocal herbivore effects on plants. We studied growth of the specialist aphid, Macrosiphoniella tanacetaria, and its host plant tansy, Tanacetum vulgare, under experimental addition of inorganic and organic fertilizer crossed with competition by goldenrod, Solidago canadensis. Because of evidence that competition by goldenrod is mediated by allelopathic compounds, we also added a treatment with activated carbon. Results showed that fertilization increased, and competition with goldenrod decreased, plant biomass, but this was likely mediated by resource competition. There was no evidence from the activated carbon treatment that allelopathy played a role which instead had a fertilizing effect. Aphid performance increased with higher plant biomass and depended on plant growth conditions, with fertilization and AC increasing, and plant competition decreasing aphid numbers. Feedbacks of aphids on plant performance interacted with plant growth conditions in complex ways depending on the relative magnitude of the effects on plant biomass and aphid numbers. In the basic fertilization treatment, tansy plants profited from increased nutrient availability by accumulating more biomass than they lost due to an increased number of aphids under fertilization. When adding additional fertilizer, aphid numbers increased so high that tansy plants suffered and showed reduced biomass compared with controls without aphids. Thus, the ecological cost of an infestation with aphids depends on the balance of effects of growth conditions on plant and herbivore performance. These results emphasize the importance to investigate both perspectives in plant herbivore interactions and characterize the effects of growth conditions on plant and herbivore performance and their respective feedbacks.

Development and reproduction of the foxglove aphid (Hemiptera: Aphididae) on invasive swallow-worts (Vincetoxicum spp.).

The foxglove aphid, Aulacorthum solani (Kaltenbach), recently was documented using the invasive species pale and black swallow-wort (Vincetoxicum rossicum (Kleopow) Barbar. and V. nigrum (L.) Moench, respectively) as host plants. Because these are new host plant records for this polyphagous species, we investigated foxglove aphid development and reproduction on pale and black swallow-wort relative to a known crop host, potato (Solanum tuberosum L.), at a 25:20°C thermophase and a photoperiod of 16:8 (L:D) h. Almost no such data have been previously reported for a noneconomic host plant. Larval development and survival, as well as adult reproductive development and fecundity, were similar between the two swallow-wort species and potato. Adult aphids lived significantly longer on pale swallow-wort than the other two host plants, but this extended longevity encompassed the postreproductive stage. Foxglove aphid population parameters were therefore similar among the three plant species as well as most previous reports on crop plants. Pale and black swallow-wort appear to be suitable secondary hosts for foxglove aphid; other factors possibly limit aphid abundance on these two plant species in the field.

Changes in aphid probing behaviour as a function of insect age and plant resistance level.

Aphids perform a series of behaviours to assess feeding suitability and, hence, to select a plant. Little information, however, is available on such behaviour after aphids have settled on a plant. Observation of probing behaviour over an extended period of time can improve our understanding of insect-plant interactions and is instrumental in the study of crop resistance. Here, we assessed the influence of aphid age and plant resistance level on aphid behaviour. An electrical penetration graph (EPG) technique was implemented to monitor the behaviour of potato aphid, Macrosiphum euphorbiae, alates on potato, Solanum tuberosum, and on both a susceptible and a resistant genotype of a wild Solanum species, S. chomatophilum. The behaviour was measured at daily intervals for the first seven days following adult emergence. The results indicated independent and interacting effects of aphid age and plant genotype on probing behaviour. Some behavioural discrepancies between susceptible and resistant genotypes were only observed after the first day, thus highlighting the limits of punctual one-day behavioural studies to assess plant resistance mechanisms. Our work supports the hypothesis that aphids continuously adapt their behaviour to the plant characteristics.

Biology and potentiality in biological control of Micraspis discolor (Fabricius) (Coleoptera: Coccinellidae).

Hosts influence on growth duration and abundance of coccinellid predators. Micraspis discolor (Fabricius) (Coleoptera: Coccinellidae) has been well known as both carnivorous and pollinivorous. Work was carried out as a series of laboratory-based influence of host species Aphis gossypii (Clover) (Hemiptera: Aphididae) and green rice leafhopper, Nephotettix sp. (Hemiptera: Cicadellidae) on M. discolors development and effect of rice growth stage on M. discolor adult preference. For field study, population of M. discolor was monitored in the rice fields throughout rice growing season for study on the relative abundance between M. discolor and hopper insects in each stage of rice growth. Results demonstrated that Nephotettix sp. was more suitably for larval growth of M. discolor than A. gossypii, with faster development time and higher larval survival. Regardless of host species, 4th larval instars were the most voracious. For feeding preference, results indicated the general preference by M. discolors adult for rice with pollen (reproductive stage) over rice without pollen (vegetative stage). In addition, female coccinellid showed feeding preference on rice pollen more than male. Consistent with the results of rice growth stage preference of adult, highest population abundance of M. discolor was found at reproductive stage of rice growth. All stages of M. discolor inhabited in rice fields, with the dominant group of adults. However, fluctuation pattern of M. discolor population synchronized with hopper insects in rice fields, population abundances of M. discolor did not related to population abundance of hopper insects.

Compatibility of garlic (Allium sativum L.) leaf agglutinin and Cry1Ac δ-endotoxin for gene pyramiding.

δ-Endotoxins produced by Bacillus thuringiensis (Bt) have been used as bio-pesticides for the control of lepidopteran insect pests. Garlic (Allium sativum L.) leaf agglutinin (ASAL), being toxic to several sap-sucking pests and some lepidopteran pests, may be a good candidate for pyramiding with δ-endotoxins in transgenic plants for enhancing the range of resistance to insect pests. Since ASAL shares the midgut receptors with Cry1Ac in Helicoverpa armigera, there is possibility of antagonism in their toxicity. Our study demonstrated that ASAL increased the toxicity of Cry1Ac against H. armigera while Cry1Ac did not alter the toxicity of ASAL against cotton aphids. The two toxins interacted and increased binding of each other to brush border membrane vesicle (BBMV) proteins and to the two important receptors, alkaline phosphatase (ALP) and aminopeptidase N (APN). The results indicated that the toxins had different binding sites on the ALP and APN but influenced mutual binding. We conclude that ASAL can be safely employed with Cry1Ac for developing transgenic crops for wider insect resistance.

[Research on population dynamics of Lonicera macranthoides aphid and natural enemy in Xiushan and evolution of pesticides].

OBJECTIVE: To study the population dynamics of aphid on Lonicera macranthoides and their natural enemy in Xiushan and control method of pesticide so as to provide scientific basis for its integrated pests management (IPM). METHOD: The field investigation and the field controlling trial were carried out for the research. RESULT: Semiaphis heraclei was the dominant species among L. macranthoides aphids. The population dynamics of apterous aphids went through five consecutive stages: initial, fluctuating, rising, peak and declining. The population dynamics of alate aphids was 4-7 days later than that apterous aphid's. Significant positive correlations were found between the population size of spiders and ladybugs which were natural enemies and number of aphids. The result of pesticides against aphids in field trial showed that 25% thiamethoxam WG, 70% imidacloprid WG and 20% acetamiprid WP had well controlling effect. CONCLUSION: Aphids on L. macranthoides could be well controlled while 25% thiamethoxam WG, 70% imidacloprid WG and 20% acetamiprid WP are sprayed during the period of aphid population raising, the early April to the mid May.

Alfalfa (Medicago sativa L.) shoot saponins: identification and bio-activity by the assessment of aphid feeding.

Biochemical components in alfalfa (Medicago sativa L.), such as saponins, can act as protecting factors against bio-stresses. Saponins are also antifeedants and show oral toxicity towards higher and lower animals. Changes in saponins, such as variation in the carbon skeleton, or hydrolysis of saponin glycosides and other conjugates, may change their biological effects. The aims of this research were to study saponin variation in different growth stages of alfalfa and to investigate the biological role of saponins in the spotted alfalfa aphid, Therioaphis maculata. Saponins from alfalfa shoots in different growth stages were extracted, chemically purified and analysed by TLC. Specific saponins such as soyasaponin1 from root and shoot and two bisdesmosides of medicagenic acid, one from shoot and another from root tissues, were identified using reference compounds allowing changes in saponin composition during plant development in different shoot tissues of alfalfa to be assessed. The response of the alfalfa aphid to feeding on alfalfa in different growth stages was studied. No significant difference in the survival of aphids, from neonate to adult, was observed, but due to the antibiotic effects of saponins, two differences were found in the onset of nymph production and cumulative nymph production. The results show that the saponin composition in alfalfa changes with plant development and this, in turn, can often negatively affect the development of specific insect pests such as the spotted alfalfa aphid, suggesting a possible biological role of alfalfa saponins.

Plant genotype mediates the effects of nutrients on aphids.

Soil nutrients, and factors which influence their concentrations and bioavailability, form a basic component of bottom-up control of ecosystem processes, including plant-herbivore interactions. Increased nutrient levels are linked, through plant defence theory, with increased levels of herbivore susceptibility. The focal point of many ecological experiments examining this link is at the species level, where the response of single species is the average of many different genotypes. Here, we focus on the genetic basis of indirect ecological interactions. We investigated the effects of nutrient concentration on the population growth of an aphid herbivore across multiple genotypes of barley in relation to plant growth rate. In general, both aphid population size and plant growth rate increased with nutrient concentration. However, they were both dependent on the interaction between nutrient concentration and barley genotype. Our data raise the testable possibility of differential defense responses between genotypes of barley, for example divergent, fixed and inducible defences against aphids. We provide evidence that the indirect effects of soil nutrients on aphid population size are mediated by the genetics of the host plant.

Homeopathic preparations to control the rosy apple aphid (Dysaphis plantaginea Pass.).

A laboratory model system with the rosy apple aphid (Dysaphis plantaginea Pass.) on apple seedlings was developed to study the effects of homeopathic preparations on this apple pest. The assessment included the substance Lycopodium clavatum and a nosode of the rosy apple aphid. Each preparation was applied on the substrate surface as aqueous solution of granules (6c, 15c, or 30c). Controls were aqueous solutions of placebo granules or pure water. In eight independent, randomized, and blinded experiments under standardized conditions in growth chambers, the development of aphids on treated and untreated apple seedlings was observed over 17 days, each. Six experiments were determined to assess the effects of a strict therapeutic treatment; two experiments were designed to determine the effects of a combined preventative and therapeutic treatment. After application of the preparations, the number of juvenile offspring and the damage on apple seedlings were assessed after 7 and 17 days, respectively. In addition, after 17 days, the seedling weight was measured. In the final evaluation of the six strictly therapeutic trials after 17 days, the number of juvenile offspring was reduced after application of L. clavatum 15c (-17%, p = 0.002) and nosode 6c (-14%, p = 0.02) compared to the pure water control. No significant effects were observed for leaf damage or fresh weight for any application. In the two experiments with combined preventative and therapeutic treatment, no significant effects were observed in any measured parameter. Homeopathic remedies may be effective in plant-pest systems. The magnitude of observed effects seems to be larger than in models with healthy plants, which renders plant-pest systems promising candidates for homeopathic basic research. For successful application in agriculture, however, the effect is not yet sufficient. This calls for further optimization concerning homeopathic remedy selection, potency level, dosage, and application routes.

Melanogenesis inhibition by gallotannins from Chinese galls in B16 mouse melanoma cells.

Nowadays herbal medicines of skin-whitening cosmetics are popular with women. We attempted to find the whitening activity compounds present in many herbal medicines used for this purpose and discuss their mechanisms in melanin biosynthesis. The 70% acetone extracts of 10 kinds of herbs were investigated for their mushroom tyrosinase activity inhibition. Among these 10 extracts, Chinese galls showed inhibitory activity against tyrosinase, with a 50% inhibitory concentration (IC(50)) value of 22 microg/ml. In a B16 mouse melanoma cell culture assay, Chinese galls dose-dependently inhibited melanin biosynthesis. Using ultraviolet A (UVA) or alpha-melanocyte-stimulating hormone (alpha-MSH) to stimulate B16 cells after Chinese gall treatment, the melanin biosynthesis of B16 cells was inhibited in a dose-dependent manner. The active compounds of Chinese galls were isolated by column chromatography, and the melanin biosynthesis inhibition in B16 melanoma cells was measured. Three gallotannins, 2,3,4,6-tetra-O-galloyl-D-glucopyranose, 1,2,3,6-tetra-O-galloyl-beta-D-glucopyranose, and 1,2,3,4,6-penta-O-galloyl-beta-D-glucopyranose, were isolated from Chinese gall extract, and their IC(50) values of tyrosinase inhibition activity were 54, 30, and 15 muM, respectively. By the mushroom tyrosinase inhibition kinetics assay, the three gallotannins were all determined to be non-competitive inhibitors. These results indicated that Chinese galls inhibit melanin biosynthesis, associated with hyperpigmentation and can be used as skin-whitening cosmetics for skin care.