Effects of insecticide acetamiprid on photosystem II (PSII) activity of Synechocystis sp. (FACHB-898)

Effects of insecticide acetamiprid on photosystem II (PSII) activity of Synechocystis sp. were investigated by a variety of in vivo chlorophyll fluorescence tests. Acetamiprid exposure increased the proportion of inactivated PSII reactive centers (PSII_X) and led to loss of active centers (PSII_A). High concentration (1.0 mM) acetamiprid decreased amplitude of the fast phase and increased the slow phase of fluorescence decay during reoxidation. The electron transport after Q_A was hindered by high concentration acetamiprid and more Q_A had to be reoxidized through S₂(Q_AQ_B)⁻ charge recombination. Acetamiprid decreased the density of the active reaction centers, electron transport flux per cross section and the performance of PSII activity but had little effect on dissipated energy flux per reaction center, antenna size and the maximum quantum yield for primary photochemistry (F_v/F_m). The target site of acetamiprid toxicity to the PSII of Synechocystis sp. was electron transfer on the acceptor side.     

Effects of fungicides JS399-19, azoxystrobin, tebuconazloe, and carbendazim on the physiological and biochemical indices and grain yield of winter wheat

The impact of four modern fungicides JS399-19 (2-cyano-3-amino-3-phenylancryic acetate) (novel fungicide), azoxystrobin (a strobilurin), tebuconazole (a triazole) and carbendazim (a benzimidazole), applied as foliar spray at the recommended field rate, on the physiology and biochemistry of the senescence process and grain yield was studied in winter wheat (Triticum aestivum L. cv. ‘Nannong No. 9918’) under natural environmental conditions. Fungicide treatments to wheat plants at growth stage [ZGS] 57 (3/4 of head emerged) significantly increased the chlorophyll (CHL) and soluble protein (SP) content and decreased the malondialdehyde (MDA) content and electrolyte leakage. Additionally, activities of the antioxidative enzymes superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in flag leaves of the fungicide-treated plants were also higher than that in untreated plants. These coincided with elevated levels of H₂O₂ and reduced level of in the fungicide-treated plants. The results suggested that the fungicide-induced delay of senescence was due to an enhanced antioxidant enzyme activity protecting the plants from harmful active oxygen species (AOS). Because all fungicides can induce the delay of wheat senescence, fungicide-treated wheat shown higher grain yield than untreated wheat. Of all tested fungicides, JS399-19, azoxystrobin and tebuconazole showed similar effects on delaying senescence of wheat and enhancing the grain yield of wheat, but JS399-19 was more efficient in general.     

Induction of hypericins and hyperforin in Hypericum perforatum L. in response to biotic and chemical elicitors

Hypericum perforatum L. produces hyperforins, a family of antimicrobial acylphloroglucinols; and hypericins, a family of phototoxic anthraquinones exhibiting anti-microbial, anti-viral, and anti-herbivore properties in vitro. To determine whether these secondary metabolites are part of the specific plant defense systems that are mediated by methyl jasmonate or salicylic acid, we used meristem cultures to assess the effects of exposure to exogenous application of these chemical elicitors. Levels of hypericins in plant tissue increased in response to both elicitor treatments; total hypericin levels increased as much as 3.3 times control levels when treated with 200 μ methyl jasmonate for 14 days. Increased hyperforin concentrations were detected when plantlets were treated with 1 m salicylic acid or 50 μ methyl jasmonate. For assessing responses to a biotic elicitor, greenhouse-grown plant materials were inoculated with the plant pathogen, Colletotrichum gloeosporioides. Levels of hypericins increased twice as much as the control when inoculated with 1 × 10⁴ spores per ml; higher doses of spores overwhelmed the plant defenses. The elevation of hypericins and hyperforin in response to chemical and biotic elicitors suggests that these secondary metabolites are components in the inducible plant defense responses of H. perforatum.     

Activation of paraquat in the earthworm Allolobophora chlorotica is mediated by NAD(P)H-cytochrome c reductase activities

This study reports that earthworms, Allolobophora chlorotica, are capable of biotransforming paraquat, a toxic herbicide, resulting in the formation of reactive oxygen species (ROS). We found that in earthworms the reduction of paraquat is mediated by NADPH- and NADH-cytochrome c reductase activities. The formation of superoxide anion (O₂⁻) from the incubation of paraquat with the earthworm extracts was demonstrated by using both Cypridina luciferin analog (CLA) chemiluminescence and the SOD-inhibitable cytochrome c reduction reaction. In addition, in vivo exposure of earthworms to paraquat in solution (24 and 48 h) was performed to investigate whether or not the herbicide affects the levels of the NAD(P)H-cytochrome c reductase activities. Although in vitro NADPH-cytochrome c reductase reduces paraquat more easily than the NADH-dependent activity, after the in vivo exposure an increase of NADH-cytochrome c reductase activity(s) by 12% compared to control values was observed, whereas NADPH-cytochrome c reductase activity was not affected. Xanthine oxidase (XO) is an enzyme implicated in paraquat toxicity, however, no XO was detected in earthworm extracts nor hypoxanthine was a source of electrons for the herbicide reduction. For comparative reasons menadione, a redox cycling quinone, was also incubated with the earthworm extracts. It was found that the incubation of menadione with earthworm extracts formed about two times more (O₂⁻) than with paraquat. It is concluded that the exposure of paraquat to earthworms could elicit radical formation and consequently toxic effects via oxidative stress-mediated mechanisms. The reduction of paraquat by the reductases leads to the formation of paraquat radical, which reacts with molecular oxygen, accounting for the formation of superoxide anion. Further studies are required to conclude that the observed increase of NADH-cytochrome c reductase activity(s) should be used as a biomarker for paraquat exposure in earthworms.     

Inhibitors of N-linked glycosylation induce systemic acquired resistance in cucumber

Localized treatment of cucumber (Cucumis sativus L. cv. Wisonsin) cotyledons with inhibitors of N-glycosylation such as tunicamycin or amphomycin resulted in systemic acquired resistance in the first leaf to the fungal pathogen Colletotrichum lagenarium. Resistance was maximal as early as 2 days after application and best results were observed when the inhibitor was used at 100 μ . The same treatment also induced salicylic acid accumulation as well as the expression of chitinase and a PR1-like protein. The systemic effect is not caused by the transport of tunicamycin, since tunicamycin was not detected in the leaves. Within 2 h after application tunicamycin inhibited N-glycosylation, but not protein synthesis as indicated by labelling experiments. The amount of large and small subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase decreased after tunicamycin treatment and after pathogen inoculation and the expression of BiP, a protein localized in the endoplasmic reticulum was enhanced. The activation of defense reactions seems to be dependent and sensitive to N-linked glycosylation.     

Bentazone and bromoxynil induce H and H2O2 accumulation, and inhibit photosynthetic O2 evolution in Synechococcouselongatus PCC7942

Using a unicellular cyanobacterium, Synechococcouselongatus PCC7942, we have shown that cytosolic acidification, O₂⁻; H₂O₂ production and photosystem II-inactivation are the causes of cell death from bentazone/bromoxynil incubations. Butyric acid evoked solely pH lowering response and yet inhibited PS II activity indicating that herbicide-caused acidification is sufficient to kill the cyanobacterial cells, but other factors like excess H₂O₂ production due to an imbalance in the peroxide sequestration machinery might be contributory. While the activities of superoxide dismutase and pyrogallol peroxidase increased consequent to herbicide incubations and displayed oligomeric states with mobility shift, catalase and glutathione peroxidase though present remained insensitive.     

Polyphosphate kinase is essential for swarming motility, tolerance to environmental stresses, and virulence in Pseudomonas syringae pv. tabaci 6605

Polyphosphate kinase (PPK), encoded by the ppk gene, is a principal enzyme responsible for synthesis of inorganic polyphosphate (poly P) from ATP in many Gram-negative bacteria. In order to elucidate the functions of poly P in Pseudomonas syringae pv. tabaci 6605, an in-frame deletion mutant of the ppk gene (ppk) was constructed. The ppk mutant did not accumulate poly P, whereas the wild-type strain accumulated a large quantity. The mutant had reduced swarming motility, even though it retains swimming motility like the parental strain. The mutant exhibited increased sensitivity to prolonged incubation and environmental stresses, such as heat shock and oxidative stress and reduced exopolysaccharide (EPS) production compared to the wild-type. Northern blot analysis revealed that expression of the rpoS gene, encoding the stationary phase sigma factor RpoS, was reduced in ppk in the logarithmic phase, indicating that rpoS is regulated by the ppk gene. The poly P deficient mutant had significantly reduced ability to cause disease in its host tobacco plant and in planta growth of the mutant was also significantly reduced in host tobacco leaves as compared to the wild-type strain. Thus, our results suggest that poly P plays an important role in the virulence of P. syringae pv. tabaci 6605.     

Synthesis, spectral characterization and antimicrobial activity of some transition metal(II) complexes with acetone p-amino acetophenone benzoylhydrazone

Complexes of the type [M(apabh)Cl] and [M(Hapabh)(H₂O)SO₄], where M = Mn(II), Co(II), Ni(II), Cu(II) and Zn(II); Hapabh = acetone p-amino acetophenone benzoylhydrazone have been synthesized and characterized. Electronic spectra and μ_eff values suggest a square planar geometry for Co(II), Ni(II) and Cu(II) chloride complexes, whereas, octahedral geometry for the sulfato complexes. ESR data show isotropic spectra for [Cu(apabh)Cl] and axial spectra for [Cu(Hapabh)(H₂O)SO₄] and d_x²_−y² as the ground state for both Cu(II) complexes. The ligand acts as tridentate monobasic in all chloro complexes bonding through two >CN and a deprotonated enolate groups, whereas tridentate neutral in all sulfato complexes coordinating through two >CN and a >CO groups. Thermal analysis (TGA & DTA) of [Ni(apabh)Cl] complex shows a multi-step exothermic decomposition pattern. The complexes show a significant antifungal activity against Rizoctonia sp., Aspergillus sp. and Penicillium sp. and a considerably fair antibacterial activity against Pseudomonas sp. and Clostridium sp. The activity increases at higher concentration of the compound.     

Infection of Tomicus piniperda (Col., Scolytidae) with Canningia tomici sp. n. (Microsporidia, Unikaryonidae)

Canningia tomici sp. n. (Microsporidia, Unikaryonidae) infects the midgut epithelium, the gut muscules, Malpighian tubules, connective tissues, adipose tissues and the gonads of the pine shoot beetle, Tomicus piniperda (L.) (Coleoptera, Scolytidae). The infection is present in populations of Tomicus piniperda in Europe and in the United States. Uninucleate oval single spores occur in two sizes: 2.8±0.4× 1.4±0.4m and 3.8±0.3×2.0±0.2m. The polar filament of this microsporidium is fixed subapically in a flat anchoring disc. The thick posterior lamellae of the binary polaroplast are asymmetric due to the lateral fixation of the polar filament.     

Prey consumption during development as well as longevity and reproduction of Typhlodromus pyri Scheuten (Acari, Phytoseiidae) at higher temperatures in the laboratory

The predatory mite Typhlodromus pyri Scheuten (Acari, Phytoseiidae) has been reported as an important predator of the European red mite, Panonychus ulmi (Koch) (Acari, Tetranychidae) in apple culture and vineyards at below 25°C. However, sufficient biological data was lacking on its efficiency at temperatures above 25°C. Therefore, the purpose of the present laboratory work was to obtain experimental data on prey consumption during development as well as longevity and reproduction of T. pyri on apple leaf discs and in Plexiglas cells at constant temperatures of 25±2°C and 30±2°C with P. ulmi as prey.The results showed that mean daily and total prey consumption by both the nymphs and adults of T. pyri decreased significantly on both the arenas as the temperature was increased from 25°C to 30°C, whereby adult prey consumption, both mean daily and total, was higher than that of nymphs. Prey consumption by both the nymphs and adults was significantly higher in the Plexiglas cells than on the leaf discs at both temperatures. Mean total prey consumption during nymphal development was 16.1 () and 12.8 () at 25°C compared to 7.0 () and 5.8 () preys at 30°C on the apple leaf discs and 46.0 () and 38.5 () at 25°C compared to 25.2 () and 20.3 () preys at 30°C in the Plexiglas cells. Mean duration of nymphal development was similar for the two sexes at the same temperature, but it was longer at 25°C than at 30°C. It was 6.0 and 4.0days on the apple leaf discs while 7.0 and 6.0days in the Plexiglas cells at 25°C and 30°C, respectively. Mean daily and total prey consumption by both male and female adults also decreased with the increasing temperature, whereby the females consumed more than double the mean total number of prey than the males on both the arenas of observation and at both temperatures: 355.4 versus 149.7 preys at 25°C and 192.2 versus 85.6preys at 30°C on the leaf discs and 826.8 versus 374.5 preys at 25°C and 488.9 versus 187.9 preys at 30°C in the Plexiglas cells. Longevity of the females was longer than males on both arenas and at both temperatures and it was longer at 25°C than at 30°C. Mean total longevity on the apple leaf discs was 68.3 () and 50.8 () days at 25°C compared to 52.5 () and 36.8 () days at 30°C, while in the Plexiglas cells it was 91.0 () and 65.8 () days at 25°C compared to 75.3 () and 48.5 () days at 30°C. Reproduction in females also decreased significantly with increasing temperature. It decreased from 62.0 to 39.0 eggs/female on the leaf discs and 75.0 to 47.1 eggs/female in the Plexiglas cells. The females laid significantly higher numbers of eggs at both temperatures in the Plexiglas cells than on the leaf discs. Oviposition period in females was 30days at 25°C on both the arenas, while at 30°C it was 26days on the apple leaf discs and 27days in the Plexiglas cells.     

Effects of different high constant and alternating temperatures on the development and prey consumption of Dicyphus tamaninii Wagner (Heteroptera, Miridae) with Aphis gossypii Glover (Homoptera, Aphididae) as prey

The present work aimed to study the effects of high constant and alternating temperatures on the development and prey consumption by Dicyphus tamaninii Wagner (Het., Miridae) with Aphis gossypii Glover (Hom., Aphididae) as prey.Results showed that the predator was able to successfully develop and reach the adult stage, on average, after 20.1 () and 20.6 () days at a temperature of 30 ± 1 °C with A. gossypii as prey. On the other hand, it was not able to develop at 35 ± 1 °C, here all N₁ instars used at the beginning of the experiments had died even before moulting to N₂ instar. At alternating temperatures of 25/15 ± 1 °C, the predator reached the adult stage 23.7 () and 23.5 () days after egg hatching, whereas when the temperatures were increased to 35/22 ± 1 °C, it took a significantly shorter period: 20.6 () and 20.9 () days. In general, the highest mortality occurred during the first two nymphal instars. The total percentage mortality during development from N₁ to adult stage was 30 % and 100 % at the constant temperatures 30 ± 1 °C and 35 ± 1 °C, respectively. At the alternating temperatures of 25/15 ± 1 °C, it was lowest with 16.5 %, while at 35/22 ± 1 °C, it was 20 %.The results of the experiments on prey consumption showed that an individual D. tamaninii consumed during its development to adult stage at 30 ± 1 °C an average of 442.2 (), 433.6 () aphids. At the constant temperature of 35 ± 1 °C, the average daily prey consumption started with 4 aphids and decreased continuously till it approached nil on the 9th day, where all predatory nymphs died. At the alternating temperatures of 25/15 ± 1 °C, the total prey consumption during development was, on average, 359.3 () and 297.0 () aphids, while at 35/22 ± 1 °C it was 348.5 () and 334.0 () aphids. The effect of the four temperatures on the prey consumption by D. tamaninii during the first 10 days after emergence was also studied. At a constant temperature of 30 ± 1 °C, a total of 446.4 () and 372.0 () aphids were consumed, compared to 40.6 () and 39.0 () at 35 ± 1 °C. At alternating temperatures of 25/15 ± 1 °C, the predatory adult consumed, on average, a total of 267.0 () and 207.0 () aphids. Increasing the temperature to 35/22 ± 1 °C resulted in a higher prey consumption by the adult predator, reaching 351.5 () and 267.7 () aphids over the same period.In conclusion, results indicated that D. tamaninii is a promising predator of the melon aphid at a temperature spectrum ranging from 15 ± 1 °C up to 35 ± 1 °C.     

A modified, four-armed olfactometer for determining olfactory reactions of beneficial arthropods

Methods using the reactions of beneficial arthropods to stimuli of prey and habitats are becoming more and more important in the biological control of pests. Especially olfactory cues, which are measurable with so called 'olfactometers' are of great importance. In the present study, a modified four-armed airflow olfactometer is described by explaining these modifications. As an example, the results obtained for the predatory ladybeetle Coccinella septempunctata are explained in detail. Various odour sources caused evident differences in the reactions of the predator, especially comparing the reared beetles and the individuals collected in the field. The volatiles emitted by the odour source 'bean plant + aphids' were attractive to 28% of the reared and to 38% of the field-collected test individuals. These values were clearly higher than in the cases of the odour source 'aphids' with 18% for both groups. About 15% of the reared and 13% of the field-collected coccinellids reacted to the olfactometer chamber without an odour source. On the other hand, it became obvious in test series with groups of adult beetles that group tests were not so suitable because the test individuals diverted each other so that the reactions to the odours were reduced. In the case of the group tests, 68% of the reared and 60% of the field-collected individuals did not react to one of the offered odour sources. In further surveys, the most favourable pre-experiment starvation period of the tested individuals and optimal duration for a single experiment were determined. For adults of C. septempunctata, the most favourable starvation time was 6 hours and the most suitable duration of a single experiment 30 minutes. In two further experiments, it was investigated whether there were differences between males and females or between beetles of different ages. There were slight differences between males and females reacting to distinct odour sources and also in the case of coccinellids of different ages but in both cases without clear tendencies.     

Investigations on side-effects of the mixed biocide GCSC-BtA on different predators of Plutella xylostella (L.) (Lep., Plutellidae) in southeastern China

The present paper deals with laboratory studies on side-effects of the mixed biocide GCSC-BtA on Plutella xylostella (L.) (Lep., Plutellidae) and its predators, e.g. Amblyseius longispinosus (Evans) (Acari, Phytoseiidae), Erigonidium graminicola (Sundvall) (Araneae, Linyphiidae), Orius similis Zheng (Het., Anthocoridae) and Coccinella septempunctata L.(Col., Coccinellidae), in comparison to the commercial insecticides, e.g. Abamectin, Tebufenozide, Dichlorvos, Cartap and Lambda-cyhalothrin.The results showed that GCSC-BtA was highly toxic to the 3^rd instar of P. xylostella with 91.18% mortality, followed by Cartap with 84.38%, Abamectin with 78.00%, Tebufenozide with 75.57%, Lambda-cyhalothrin with 63.75% and Dichlorvos with 50.86% mortality. On the other hand, GCSC-BtA was found to be comparatively less toxic to the predators, causing 31.11%, 13.33%, 11.54% and 6.00% mortalities in A. longispinosus, E. graminicola, O. similis and C. septempunctata, respectively. For comparison, the mortalities recorded for Abamectin, Tebufenozide, Dichlorvos, Cartap, Lambda-cyhalothrin were 72.94%, 55.55%, 70.00%, 53.26% and 98.85% in A. longispinosus, 46.51%, 55.10%, 60.00%, 46.00% and 73.68% in E. graminicola, 22.00%, 16.00%, 35.71%, 26.78% and 81.03% in O. similis, 15.55%, 19.64%, 28.00%, 16.66% and 41.79% in C. septempunctata, respectively.     

Parasitoids of Saperda populnea (L.) (Coleoptera: Cerambycidae) on aspen (Populus tremula L.) in Bulgaria

The parasitoids of Saperda populnea (L.) (Col.: Cerambycidae) were studied in Bulgaria during the period 1997–2001. Galls with pest larvae were collected from young aspen trees at eight locations (Sofia, Kokaliane, Plana, Churek, Gorni Lom, Gintzi, Dolno Kamartzi and Klisura) and examined under laboratory conditions. Four species were reared from 1118 galls containing overwintering pest larvae: Iphiaulax impostor (Scop.) (Hym.: Braconidae), Dolichomitus populneus (Ratz.), Schreineria populnea (Gir.) (Hym.: Ichneumonidae) and Billaea irrorata (Meig.) (Dipt.: Tachinidae). All of the parasitoids were solitary. Schreineria populnea was found as a new parasitoid of the host in Bulgaria. B. irrorata emerged mainly from late-stage S. populnea larvae; the remaining species from both early- and mid-stage larvae. In the parasitoid complex of S. populnea, the most numerous were B. irrorata (59.8%) and D. populneus (29.4%). The percentage of host larval parasitism varied from 2.4 to 33.3, with an average of 9.7%. B. irrorata was the most important in reducing the pest number. The average mortality of S. populnea caused by the tachinid was 5.8%, and the highest mortality observed in individual study was 20.2%.