Shallot latent virus,a new carlavirus

A new carlavirus, apparently omnipresent in shallot (Allium ascalonicum) without causing symptoms, is described as shallot latent virus. It has also been detected in naturally infected onion (A. cepa) and leek (A. porrum). The virus was easily transmissible in sap and could be transferred withMyzus ascalonicus. Infection after inoculation was symptomless in onion, leek,A. fistulosum andA. jailae. Chenopodium album, C. amaranticolor andC. quinoa reacted with local lesions.A. neapolitanum andA. schoenoprasum and 20 other plant species were immune. Dilution end-point was 10⁴–10⁵, thermal inactivation at ca. 80°C and ageing in vitro 8–11 days. Purification by molecular sieving on Sephadex G-200 followed by equilibrium density-gradient centrifuging in CsCl was successful. Sedimentation coefficient was 147.5 S, bouyant density 1.313 g/cm³ and molecular mass of protein subunits 23 200 dalton. With the antiserum (titre 1024) distant serological relationships to some carlaviruses were determined. No inclusion bodies were observed with the light microscope. With the electron microscope a high concentration of straight or slightly curved particles with a normal length of 650 to 652 nm could easily be detected in crude sap and in purified preparations.     

Steinernematid and heterorhabditid nematodes for biological control of the giant looper,Boarmia selenaria

The pathogenicity of steinernematid and heterorhabditid nematodes to caterpillars of the giant looper,Boarmia selenaria (Schiffermüller) (Lepidoptera: Geometridae), was determined under laboratory conditions. High mortality (over 80%) was achieved, in a petri dish bioassay, with a minimum of 20,000 infective juveniles (IJ) ofSteinernema carpocapsae (Filipjev) (=Neoaplectana carpocapsae Weiser), strain ‘All’. The LD₅₀ was 4250 IJ per caterpillar. Similar results were obtained withS. carpocapsae, strain ‘Mexican’,Steinernema sp., strain ‘CR’;Heterorhabditis bacteriophora, strain ‘HP88’; andHeterorhabditis sp., strain ‘IS’. The first and second instars were the most susceptible stages to nematode infection. A laboratory model assay indicated that 80–90% of the caterpillars were killed when they migrated to the soil to pupate; this occurred when IJ were applied to sandy and heavy soils at a concentration of 400 IJ/cm³ soil. A greenhouse test reconfirmed the effectiveness of utilizing the entomopathogenic nematodes in an “ambush” system against caterpillars migrating to the soil.     

Laboratory evaluation of steinernematid and heterorhabditid nematodes for control of the beetleMaladera matrida

The potential use of steinernematid and heterorhabditid nematodes against the beetleMaladera matrida Argaman (Coleoptera: Scarabaeidae) was determined under laboratory and greenhouse conditions. Infective juveniles (IJ’s) of the nematodeHeterorhabditis sp., Hp88 strain, mixed with soil at a concentration as low as 50 IJ’s/cm³ soil, resulted in 86% control. No increase in control was obtained with higher nematode concentrations. Soil surface application of the nematodes at concentrations of 160 and 640 IJ’s/cm² was sufficient to obtain 87% and 86% mortality, respectively, even at a depth of 40 cm below the soil surface. TheHeterorhabditis sp. Hp88 strain was found to be the most pathogenic to the beetle grubs at 25°C. Lower control levels of 30-47% were achieved byHeterorhabditis sp. HL81 strain,S. feltiae ‘All’ strain and 5.bibionis CR strain. The nematodeHeterorhabditis sp. BS strain did not have any effect on insect mortality. However, at a temperature of 16°C, the HL81 strain ofHeterorhabditis sp. was the most effective. Grubs 3-5 wk old were found to be the most susceptible developmental stage of infection ofHeterorhabditis sp. Hp88 strain. Pupae, which were exposed to the nematodes in the same experimental regime, were not affected. The results obtained in the present study suggest thatM. matrida is an attractive candidate for biological control by entomoparasitic nematodes.     

Effect of avermectin B1 on the development and fecundity of the california red scale,Aonidiella auranth,and on two of its natural enemies:Chilocorus bipustulatus andAphytis holoxanthus

Various immature stages of male and female California red scale,Aonidiella aurantii (Maskell) (Homoptera: Diaspididae), were treated with avermectin B₁ (MK-936) at concentrations ranging from 10 to 100 ppm. Treatments at 10 ppm completely arrested crawler development and 20 and 100 ppm caused 100% mortality of 48-h and 7-day-old male and female 1st instar nymphs, respectively. The development of the 2nd instar was slightly affected by MK-936 at up to 40 ppm. Sublethal doses of the chemical applied to immature stages caused no dramatic reduction in the fecundity of the subsequent, mature females. MK-936 applied to male prepupal and pupal stages and mated females did not prevent male emergence, nor did it affect the ability of males to copulate and inseminate female scales, or the female fecundity. The development and adult emergence of treatedAphytis holoxanthus DeBach (Hymenoptera: Aphelinidae) larvae and pupae parasitizingChrysomphalus aonidum L. scales were not influenced by a 9 ppm MK-936 treatment; leaves sprayed with this concentration had residual toxicity onA. holoxanthus adults for only 24 h post-treatment. Feeding byChilocorus bipustulatus L. (Coleoptera: Coc-cinellidae) on 0.9 ppm MK-936-treated Florida red scales resulted in 100% mortality of 2nd instars and induced sterility in the females.     

Management of wheat blast with synthetic fungicides,partial resistance and silicate and phosphite minerals

Field trials conducted on a yellow-red latossol (pH 6.0), replicated in 2010 and 2011, sought to examine the effect of silicon, phosphite minerals, synthetic fungicides and genetic resistance for wheat blast management (Magnaporthe grisea) in Central Brazil. Disease intensity was measured on cvs. BRS 264 and BR18 subjected to the following Si treatments: pre-plant furrow application of Ca & Mg silicate (300 kg ha^-1); post-plant scattered application of Ca & Mg silicate on top of the soil (1 ton ha^-1); multiple foliar SiO₂ applications (30 g l ^-1); and non-treated control. Blast incidence and severity were scored. Further experiments were conducted on cv. BR-264, for examination of the effect of potassium phosphite and synthetic fungicides on wheat blast intensity, with the following treatments: K₂HPO₃ (1ml l ^-1); epoxinazole + pyraclostrobin (700 ml ha^-1); tebuconazole (600 ml ha^-1); tebuconazole + trifloxystrobin (750 ml ha^-1); and non-treated control. In 2010, disease intensity was lower than in 2011. In the silicate experiments, disease was significantly lower when plants were treated with foliar or furrow silicate. Si applications significantly reduced disease in BRS-264. While BR-18 consistently demonstrated lower disease levels, cv. BRS-264 generally responded more markedly to silicon applications. In the phosphite/fungicide experiment of 2010, all treatments reduced disease when compared with the control, and in 2011 phosphite efficiency was not significantly different from some fungicide treatments. Synthetic fungicides demonstrated an average blast control of 55% by severity values. Yields were increased in the phosphite-treated plots (by 9–80%), in the Si treatments (by 26–92%), and more so, and more consistently, with synthetic fungicides (by 90–121%). Combined results of all field studies, carried out under environmental conditions highly conducive to disease, indicated that control of wheat blast necessitates the joint integration of several alternatives for efficient disease management.     

Comparative toxicity of several insecticides to eggs,larvae and adults of the Egyptian cottonworm,Spodoptera littoralis,in laboratory trials

Insecticides commonly used in cotton fields in Israel against lepidopterous pests were tested against eggs, as well as against 2nd-instar larvae and adults raised from eggs ofSpodoptera littoralis (Boisduval) collected in cotton fields in the Bet She’an Valley. Methomyl, chlorpyrifos, methidathion, monocrotophos, ethyl parathion, and methyl parathion were effective against eggs even at low doses, profenofos and phosfolan were less active, and azinphos-methyl was ineffective. The doses needed for 90% kill (LD ₉₀ ) of the 2nd-instar larvae were 8.5, 35, 280, 1300 and 3400 g a.i./1000 m ² for chlorpyrifos, methomyl, profenofos, methyl parathion and ethyl parathion, respectively; monocrotophos was inactive against 2nd-instar larvae even at relatively high doses. The LD ₉₀ of adults was reached with 16, 32, 1700 and 6100 ga.i./l000 m² of chlorpyrifos, methomyl, ethyl parathion and profenofos, respectively. Only chlorpyrifos and methomyl gave successful control of all three stages of the insect tested, at doses close to those generally used with aerial applications.     

Integrative diagnosis and molecular phylogeny of dagger and needle nematodes of olives and grapevines in the island of Crete,Greece, with description of Xiphinema cretense n. sp. (Nematoda,Longidoridae)

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated and wild olive and grapevine in Crete Island were investigated. Morphological and morphometrical studies identified five Longidorus and six Xiphinema species, with frequencies of prevalence (for wild and cultivated olives and grapevines, respectively) as follows: Longidorus closelongatus (2.0–13.3 %), L. cretensis (1.0–6.7 %), L. moesicus (13.3 % only in grapevines), L. orientalis (3.3 % only in grapevines), L. pseudoelongatus (7.0 % only in olives), Xiphinema cretense n. sp. (3.0 % only in olives), X. index (3.0–23.3 %), X. israeliae (6.3 % only in olives), X. italiae (3.3–10.0 %), X. pachtaicum (26.7–42 %) and X. simile (3.3 % only in grapevines). Xiphinema cretense n. sp. is characterized by a body size 3,872–6,135 μm long, lip region anteriorly rounded, separated from the rest of the body by a depression, odontostyle and odontophore 140.6 and 80.3 μm long respectively, vulva position at 46.0–50.5 %, female tail 31.0–38.0 μm long, nearly hemispherical with curvature essentially dorsal and with a tip completely rounded or presenting a very short bulge, c ratio (119.1–186.9), c’ ratio (0.7–0.8). Molecular characterisation using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids.     

Pyrethroid synergism and prevention of emergence inTribolium castaneum andMusca domestica vicina by the insect growth regulator RO 13-5223

Ethyl [2-(4-phenoxyphenoxy)ethyl] carbamate (RO 13-5223) at a dietary concentration of 100 mg kg^-1 synergized the toxicity of thetrans- andcis-isomers of permethrin and cypermethrin in inhibiting the growth (measured as gain in larval weight) ofTribolium castaneum andMusca domestica vicina. With both species the synergism factor forcis-cypermethrin with 100 mg kg^-1 synergist was 1.5- to twofold for RO 13-5223 and about fourfold for piperonyl butoxide. Synergism was more pronounced with first instar than with fourth instarT. castaneum larvae. Methoprene was not a pyrethroid synergist withT. castaneum larvae, so the synergistic effect of RO 13-5223 appears to depend on its structural features and not its insect-growth-regulator activity. Joint application of RO 13-5223 and pyrethroids resulted in a dual effect on bothT. castaneum andM. domestica: increased inhibition of larval growth due to pyrethroid synergism, and progeny suppression — expressed by larval and pupal mortality — due to RO 13-5223 juvenilizing activity.     

In planta detection of Macrophomina phaseolina from jute (Corchorus olitorius) by a sodium acetate-based direct PCR method

Macrophomina phaseolina (Tassi) Goid. is the most important pathogen of jute and primarily causes seedling blight, leaf spot and stem rot. The pathogen was detected from field samples by a simple method of direct PCR (dPCR) which obviates the steps of DNA extraction. The leaf bits were treated with a lysis buffer at 65°C for 25 min, whereas the stem pieces were initially incubated at 65°C for 5 min followed by incubation at 60°C for 25 min and the lysate was used as PCR template. Based on the type of tissue, the composition and concentration of lysis buffer systems were optimized. For leaf samples the optimized buffer system composed of 20 mmol l ^-1 tris (hydroxymethyl aminomethane (Tris)-Cl (pH 8.0), 1.5 mmol l ^-1 ethylene diamine tetra acetate (EDTA) (pH 8.0), 1.4 mol l ^-1 sodium acetate and 200 μg/ml proteinase K. Further, 3% PVP (w/v) and β-mercaptoethanol (1% w/v) were added into the buffer. In case of stem samples, PVP was not applied and higher concentrations were used for other components. M. phaseolina could be detected from both leaf and stem samples generating amplicon of 350 bp. This is the first report of detecting M. phaseolina by a direct PCR method without DNA extraction.     

Insecticidal,repellent and fungicidal properties of novel trifluoromethylphenyl amides

Twenty trifluoromethylphenyl amides were synthesized and evaluated as fungicides and as mosquito toxicants and repellents. Against Aedes aegypti larvae, N-(2,6-dichloro-4-(trifluoromethyl)phenyl)-3,5-dinitrobenzamide (1e) was the most toxic compound (24 h LC₅₀ 1940 nM), while against adults N-(2,6-dichloro-4-(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (1c) was most active (24 h LD₅₀ 19.182 nM, 0.5 μL/insect). However, the 24 h LC₅₀ and LD₅₀ values of fipronil against Ae. aegypti larvae and adults were significantly lower: 13.55 nM and 0.787 × 10⁻⁴ nM, respectively. Compound 1c was also active against Drosophila melanogaster adults with 24 h LC₅₀ values of 5.6 and 4.9 μg/cm² for the Oregon-R and 1675 strains, respectively. Fipronil had LC₅₀ values of 0.004 and 0.017 μg/cm² against the two strains of D. melanogaster, respectively. In repellency bioassays against female Ae. aegypti, 2,2,2-trifluoro-N-(2-(trifluoromethyl)phenyl)acetamide (4c) had the highest repellent potency with a minimum effective dosage (MED) of 0.039 μmol/cm² compared to DEET (MED of 0.091 μmol/cm²). Compound N-(2-(trifluoromethyl)phenyl)hexanamide (4a) had an MED of 0.091 μmol/cm² which was comparable to DEET. Compound 4c was the most potent fungicide against Phomopsis obscurans. Several trends were discerned between the structural configuration of these molecules and the effect of structural changes on toxicity and repellency. Para- or meta- trifluoromethylphenyl amides with an aromatic ring attached to the carbonyl carbon showed higher toxicity against Ae. aegypti larvae, than ortho- trifluoromethylphenyl amides. Ortho- trifluoromethylphenyl amides with trifluoromethyl or alkyl group attached to the carbonyl carbon produced higher repellent activity against female Ae. aegypti and Anopheles albimanus than meta- or para- trifluoromethylphenyl amides. The presence of 2,6-dichloro- substitution on the phenyl ring of the amide had an influence on larvicidal and repellent activity of para- trifluoromethylphenyl amides.     

Ulvan effect on conidial germination and appressoria formation of Colletotrichum gloeosporioides

Ulvan is an algal polysaccharide known for its ability to induce resistance to plant diseases such as the Glomerella leaf spot of apple caused by Colletotrichum gloeosporiodes. This study was aimed at investigating microscopically, in tests in vitro and in vivo, whether ulvan interferes in the development of pre-infective structures of C. gloeosporioides. Conidial germination and appressoria formation were monitored hourly on agar and cellophane, and at 48 h on water- and ulvan-treated susceptible as well as resistant apple leaves. Amendment of agar with ulvan (10 mg ml^?1) enhanced the germination and resulted in longer germ tubes at 7 h of incubation. On cellophane it significantly delayed appressoria formation up to 8 h, but later after 14 h increased the number of appressoria per conidium. Spraying of susceptible leaves with ulvan 6 days before inoculation decreased disease severity by 50%. This was associated with inhibition of appressoria formation and stimulus in growth of germ tubes, without interfering with conidial germination, when compared with both water-treated control and resistant plants. Appressorium formation occurred preferentially on anticlinal walls of epidermal cells and its location was not influenced by host resistance or by ulvan treatment. This study suggests a new mode of action for ulvan interfering with appressorium formation that could protect apple plants against C. gloeosporioides infection.     

Synthetic pyrethroids and avermectin for controlling the grapevine pestsLobesia botrana,Cryptoblabes gnidiella andDrosophila melanogaster

Field treatments in a vineyard with 0.015 or 0.01% a.i. of cypermethrin, fenvalerate, fenpropathrin or AC-222,705 were more efficient in controlling the grape-berry moth (Lobesia botrana Schiff.) and the honeydew moth (Cryptoblabes gnidiella Mill.) than four standard treatments consisting of two with 0.05% a.i. fenitrothion and two with 0.075% a.i. diazinon. In pyrethroid-treated plots, infestation at the end of the trials ranged between 2.5 and 12%, compared with 21% in the standard treatment plots and 34% in the untreated plots. Cypermethrin, fenpropathrin and AC-222,705 exhibited similar field activity, while that of fenvalerate was somewhat lower. Under laboratory conditions, cypermethrin at 0.005 and 0.01% a.i. was significantly more potent than fenvalerate, fenpropathrin and AC-222,705; at a higher concentration, 0.015% a.i., all pyrethroids were highly active, with mortality ranging between 75 and 95%. Under laboratory conditions the vinegar fly (Drosophila melanogaster Meig.) was in general more susceptible to pyrethroids than was the grape-berry moth. Cypermethrin and AC-222,705 at 0.005% a.i. and avermectin at 0.0035% a.i. were potent compounds against the vinegar fly and more active than fenvalerate and fenpropathrin.     

Development of conventional and real-time PCR protocols for specific and sensitive detection of Colletotrichum capsici in chilli (Capsicum annuum L.)

Anthracnose, caused by Colletotrichum capsici, is a major disease of chilli (Capsicum annuum L.) affecting both fruit and seed quality. The pathogen is both internally and externally seedborne. However, a rapid and sensitive method for detection of this pathogen in seeds is currently limited. In this study, a polymerase chain reaction (PCR) method based on sequence characterized amplified region (SCAR) marker was developed for specific and sensitive detection of C. capsici in chilli seeds and fruits. The developed SCAR primers were highly specific to C. capsici and resulted in the amplification of an expected 250-bp fragment from genomic DNA of all seven of the C. capsici isolates tested. No amplification occurred when the SCAR primers were tested with genomic DNA from three other fungal isolates and four other Colletotrichum species. The SCAR primers successfully amplified similar sized fragments from DNA derived from C. capsici-infected chilli fruits. The molecular detection sensitivity of C. capsici was 1 pg of purified C. capsici DNA template and 25 ng of DNA from C. capsici-infected chilli fruits. A real-time PCR assay was also developed using SYBR Green chemistry for detection of C. capsici in chilli fruits and seeds. The standard curve obtained showed a linear correlation between copy number of the cloned target DNA sequence of C. capsici and cycle threshold (Ct) values, with R² of 0.98. These PCR-based assays may be highly useful in detection of this important pathogen in chilli seeds and fruits in plant quarantine laboratories.     

Potential use of Eucalyptus globulus essential oil against Phytophthora colocasiae the causal agent of taro leaf blight

This study was carried out to determine the antimicrobial activity of essential oil derived from leaves of eucalyptus (Eucalyptus globulus) against Phytophthora colocasiae, the causal agent of taro leaf blight (TLB). Essential oil was obtained by Clevenger-type water distillation. The major compounds in this essential oil were identified using gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC/MS). Antimicrobial activity of the essential oil was tested in vitro against mycelial growth, sporangia and zoospores germination of P. colocasiae. Additionally, in situ tests were conducted on detached healthy taro leaves discs and evaluating necrosis symptoms of TLB were assessed. Results of the chemical composition of the essential oil analysis showed that, 1.8-cineole (26.4 %), α-pinene (14.1 %) and p-cymene (10.2 %) are the most abundant compounds. In addition, four components could be identified in noticeable amounts (18.1 % in the total): regulator G1/G2 and its derivatives as well as flavesone, a regulator G3 derivative. G is a generic term for a family of 2,3-dioxabicyclo[4,4,0] decane system growth regulators. Here, G-regulators and derivatives are described for the first time in E. globulus. Antimicrobial activity of the essential oil was recorded with total inhibition of mycelia growth and sporangia germination at 0.625 mg/ml, while the complete inhibition of zoospores germination was recorded at 0.156 mg/ml. In situ results showed that essential oil completely inhibited the appearance of disease symptoms, necrosis development and sporulation at 3.5 mg/ml. These results demonstrated that, the essential oil of Eucalyptus globulus has high ability to inhibit the development of P. colocasiae, and might be used for controlling TLB.     

Plant ferredoxin-like protein enhances resistance to bacterial soft rot disease through PAMP-triggered immunity in Arabidopsis thaliana

The protection of plants against bacterial disease is one of the important issues that need to be studied in agricultural applications. The application of a transgene, such as a gene that encodes plant ferredoxin-like protein (PFLP), to generate resistant plants is one possible strategy. Our previous reports have demonstrated that transgenic plants that express extracellular PFLP (ESF plants) are more resistant to bacterial pathogens. This protein intensifies the hypersensitive response (HR) in plants when they are infiltrated by a pathogen-associated molecular pattern (PAMP), harpin (HrpZ), from Pseudomonas syringae. In addition, this intensified HR is associated with the expression of membrane-bound NADPH oxidase. Thus, we attempted to determine the involvement of PFLP in intensifying PAMP-triggered immunity (PTI) to enhance disease resistance. First, we showed that transgenic Arabidopsis plants with the pflp gene were resistant to bacterial soft rot caused by Pectobacterium carotovorum subsp. carotovorum (Pcc). Then, the fliC gene which encoded flagellin from Pcc was cloned and expressed. The FliC protein was used in the functional study with PFLP in Arabidopsis Col-0 plants. The reactive oxygen species (ROS) generation and HR ratio were induced by the treatment with both PFLP and FliC together, but they were not induced by treatment with PFLP or FliC alone. Similar results were confirmed in ESF plants, where FliC elicited rapid ROS accumulation and callose deposition. Moreover, we demonstrated that the PFLP-intensified ROS generation and HR were related to Ca²⁺ influx and activation of NADPH oxidase. We concluded that the PFLP-intensified disease resistance is associated with the intensification of PAMP-triggered immunity.     

Further characterization of dandelion yellow mosaic virus from lettuce and dandelion

A damaging virus isolated in the Netherlands from lettuce was studied and compared with a virus isolated from dandelion orginating from Czechoslovakia. It was found to biologically resemble dandelion yellow mosaic virus incompletely described from dandelion and lettuce in Great Britain (Kassanis, 1944, 1947) and from dandelion in Germany (Hein, 1963). Mechanical transmission was greatly improved by buffer solution and transmission byMyzus persicae seemed to be in the non-persistent manner. Longevity in vitro of the virus hardly exceeded one day. Thermal inactivation was between 60 and 65 °C and the dilution end-point was between 10 000 and 100 000. It was still infectious in leaf material dried and stored over CaCl₂ at 4 °C for 6 1/2 years. The virus was isolated and purified with difficulty and was found to consist of one type of spherical particle of ca 30 nm diameter, with a sedimentation coefficient of 159 S, a buoyant density of 1.42 g.cm^?3 and an A₂₆₀/A₂₈₀ ratio of 1.67. An antiserum was prepared with a titre of 256 in the agar double-diffusion test. The virus could be identified in crude extracts from lettuce andChenopodium amaranticolor by enzyme-linked immunosorbent assay (ELISA), but not by agar double diffusion. It could only be visualized in crude sap in the electron microscope after trapping of virus particles on antiserum-coated grids. The virus cannot yet be assigned to any known virus group. It is of potential economic importance to lettuce because of its occurrence in widely differing regions in Europe, its aggressiveness and virulence on 22 out of 23 lettuce cultivars tested (and on endive) and its pathogenicity toLactuca genotypes which are resistant to lettuce mosaic virus and other important pathogens of lettuce. ‘Laibacher Eis’ was the only cultivar showing some tolerance.     

Resistance mechanisms and risk assessment regarding indoxacarb in the beet armyworm,Spodoptera exigua

The beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), is a worldwide polyphagous pest with a strong ability to develop resistance. After 12 generations of selection by indoxacarb, a resistant strain (RR-indox) was obtained with a 240.04-fold resistance ratio compared with the susceptible strain (WH-SS), which was derived from the Wuhan Institute of Vegetable Science. The LC₅₀ for the susceptible strain was 0.23 mg l ^-1 and it increased to 55.21 mg l ^-1 after 12 selected generations. The estimated realized heritability (h²) of the RR-indox strain was 0.25. The number of generations required for a 100-fold increase in LC₅₀ was estimated to be nine under the 90% lethal dose. The inhibitors piperonyl butoxide (PBO), triphenyl phosphate (TPP), and diethylmeleate (DEM) had synergistic ratios of 0.87-, 0.31- and 0.36-fold in RR-indox, and 0.51-, 0.09- and 0.12-fold in WH-SS, respectively. A 3.75-fold increased level of activity of glutathione S-transferase was found in RR-indox compared with WH-SS, whereas a 1.99-fold higher activity level of carboxylesterase was observed. No significant differences in the cuticular penetration rate were found between RR-indox and WH-SS. A DNA fragment of 257 bp length of the IIS6 region of the voltage-gated sodium channel gene was amplified and sequenced from both RR-indox and WH-SS. An amino acid substitution from leucine (CTT) in WH-SS to phenylalanine (TTT) in RR-indox was observed at site 1014. These results show the resistance mechanisms of S. exigua to indoxacarb to be associated with glutathione S-transferase, carboxylesterase and a Leu-1014-Phe mutation.     

Response ofSpodoptera littoralis (Boisd.) andEarias insulana (Boisd.) larvae to azadirachtin and salannin

The effect of azadirachtin and salannin, two triterpenoids isolated from seeds of neem (Azadirachta indica A. Juss), on the feeding response ofSpodoptera littoralis (Boisd.) andEarias insulana (Boisd.) larvae, was investigated. Styropor (foamed polystyrene) lamellae were painted on both sides with a mixture of 5% sucrose with different concentrations of either azadirachtin or salannin dissoled in methanol-water (3∶7). Azadirachtin strongly suppressed feeding inS. littoralis larvae even at 0.001%, whereas salannin showed some antifeedant activity at 0.005% and above. Larvae ofE. insulana were deterred from feeding on azadirachtin-treated lamellae even at 0.005%, whereas salannin was effective only at 0.01% and above. Azadirachtin applied on cotton leaves deterred larvae ofS. littoralis from feeding at all concentrations ranging between 0.001 and 0.02%.     

Residue analysis of butocarboxim and aldicarb applied in a drip-irrigated peach grove by the HPLC post-column fluorogenic labeling technique

The new high-performance liquid chromatography (HPLC) post-column fluorogenic labeling technique was used for the analysis of residues of butocarboxim, aldicarb and their sulfoxide and sulfone metabolites in peach leaves and fruits. In order to simplify the procedure, the different metabolites were separated by column chromatography during the cleanup process and the residues were determined by HPLC employing an aqueous methanol solvent in the isocratic mode. The limit of detection for the two insecticides was 0.1 μg/g in the leaves and 0.02 μg/g in the fruit. Aldicarb (10% granular formulation) and butocarboxim (50% E.C. diluted 1:5 with water) were applied to the soil in a drip-irrigated peach grove, by placement under one dripper on one side of each tree. The insecticides were taken up by the tree and distributed almost equally on both sides of the tree. The concentration of the insecticides was somewhat lower in the younger leaves. Residue levels of 0.4 to 0.9 μg/g were detected in the fruits at harvest, 10 and 18 days after the last treatment. Based on the results from the field experiment it was concluded that one of the advantages of drip irrigation was that it enabled adequate control of the green leafhopper (Asymmetrasca decedens) throughout the summer with relatively low dosages of systemic insecticides.     

Alien scale insects (Hemiptera Coccoidea) in European and Mediterranean countries: the fate of new and old introductions

This contribution focuses on recent interceptions and introductions of alien scale insects and their current distribution in European and Mediterranean countries. Data and collections were gathered in markets, nurseries, and botanical gardens, mostly in Italy, either indoors or outdoors. New or recent records of the following alien species are presented: Exallomochlus hispidus (Morrison); Ferrisia virgata (Cockerell) (Pseudococcidae); Coccus viridis (Green); Milviscutulus mangiferae (Green) (Coccidae); Aonidiella orientalis (Newstead); Aspidiotus destructor Signoret; Aulacaspis tubercularis Newstead; Fiorinia fioriniae Targioni Tozzetti; Lepidosaphes pinnaeformis (Bouché); Pseudaulacaspis brimblecombei Williams (Diaspididae). New data and pest status of Phoenicococcus marlatti Cockerell (Phoenicococcidae) and Trabutina mannipara (Hemprich & Ehrenberg) (Pseudococcidae) are also reported. The possible repeated introductions of the latter from North Africa to south Italy by trans-Mediterranean winds, is hypothesized.