Laboratory evaluation of diatomaceous earth deposits mined from several locations in central and southeastern Europe as potential protectants against coleopteran grain pests

Diatomaceous earth (DE) deposits from regions of central and southeastern Europe were evaluated for their insecticidal efficacy against Sitophilus oryzae (L.) (Coleoptera: Curculionidae), Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) and Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) in comparison with the commercially available DE formulation SilicoSec. The effects of temperature, RH, grain commodity (wheat, barley, maize, rice), application method (spraying vs. dusting) were evaluated. FYROM, a DE from the Former Yugoslavian Republic of Macedonia, was the most effective of the DE deposits for grain treatment, whereas the least effective was from Greece (named Crete). However, for surface treatment, Slovenia was the most effective followed by Elassona 1 and Begora. Increase of temperature increased DE efficacy, while the reverse was noted with the increase of RH. Furthermore, the DEs were more effective in barley or wheat than in maize or rice. Neither the mined DEs nor SilicoSec were able to suppress progeny production of the tested species after previous exposure on the treated commodities. Generally, dust application of DEs was more efficacious than spraying against S. oryzae and T. confusum. However, spraying of wheat significantly reduced the bulk density (test weight) compared to dusting. For surface treatment, after 1 d of exposure, Slovenia was the most effective of the mined DEs followed by Elassona 1 and Begora, whereas after 6 d of exposure the mortality was almost complete (>99%) with all three DEs. More than 6 d of exposure were required for an effective control of T. confusum adults with the remainder of the mined DEs.     

Insecticidal efficacy of two pirimiphos-methyl formulations for the control of three stored-product beetle species: Effect of commodity

Laboratory bioassays were conducted to evaluate the insecticidal effect of two pirimiphos-methyl formulations, an emulsifiable concentrate (EC) and a capsule suspension (CS) against adults of Sitophilus oryzae, Rhyzopertha dominica and Tribolium confusum in wheat, maize and rice. Pirimiphos-methyl was applied at two dose rates, 2 and 4 ppm, and adult mortality was measured after 2, 7, 14 and 21 days of exposure in the treated commodity. Progeny production was assessed 65 d after the removal of the parental adults. S. oryzae adult survival was significantly higher in rice than in wheat and maize, whereas T. confusum mortality was significantly higher in maize than in the other grains. From the species tested, the most susceptible species was S. oryzae, for which mortality reached 100% after 7 days of exposure in treated wheat and maize, followed by T. confusum and R. dominica. Progeny production was significantly suppressed by pirimiphos-methyl in the case of S. oryzae, but not in the case of R. dominica. In general, only few differences in performance between the EC and the CS formulation were detected. We conclude that the type of grain commodity significantly affects mortality after the application of the two pirimiphos-methyl formulations tested, but this effect is species-dependent.     

Persistence and efficacy of spinosad on wheat,maize and barley grains against four major stored product pests

The insecticidal and residual effect of spinosad on wheat, maize and barley grain was evaluated in the laboratory against adults of Sitophilus oryzae (F.) (Coleoptera: Curculionidae), Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), Tribolium confusum (DuVal) (Coleoptera: Tenebrionidae), Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae) as well as against larvae of T. confusum. Spinosad was applied as a solution to 2 kg lots of each commodity at three concentrations, 0.1, 0.5 and 1 ppm, and the treated grain quantities were kept at 25 °C and 65% RH. Samples were taken from each concentration-commodity combination at the day of storage and every 30 d for 6 consecutive months (6 bioassays). The test species were exposed for 14 d to the samples and mortality and reproduction were assessed over this exposure interval. With the exception of T. confusum, 1 ppm of spinosad was highly effective against the remainder of the tested species and provided protection for a period of storage at least 4 months. Although in general, spinosad performance was not very much affected by the grain type, efficacy on maize was less stable over the 6-month period of storage and declined sooner compared to the other commodities. Spinosad almost suppressed progeny production of R. dominica during the storage period, but did not suppress progeny of the other species, since progeny were recorded even 30 d post application especially with the lowest of the tested concentrations. The results of this study indicated that spinosad may provide suitable protection for 6 months against S. oryzae or R. dominica, but is not suitable for long-term protection against T. confusum or C. ferrugineus.     

Time-mortality relationships of larvae and adults of grain beetles exposed to extreme cold

Mortality effects of low temperature exposure to −16 °C were investigated on larvae and adults of the grain beetles Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae), Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae) and Trogoderma granarium (Everts) (Coleoptera: Dermestidae), in the laboratory. The main effects and interactions of exposure time (0.2, 0.5, 1, 2, 4, 8, 12, 24 and 48 h), developmental stage (larva and adult) and individual’s age (young and old) were examined. After 4 h of exposure 100% mortality has been achieved in most of the treatments. The only exceptions were that of old larvae of T. granarium and O. surinamensis, and adults of T. granarium which needed 48, 8, and 24 h, respectively. T. granarium was the most cold-hardy species irrespective of developmental stage, and age, followed in decreasing order by O. surinamensis and T. confusum. Younger adults were generally more susceptible to cold in the cases of O. surinamensis and T. confusum. On the contrary, older adults of T. granarium suffered higher mortality than younger ones. Larvae of T. granarium were generally more cold-tolerant than adults but the opposite pattern was observed in O. surinamensis and T. confusum. Main effects of exposure time, developmental stage and individual’s age on mortality proved to be significant for all species, with the exception of T. granarium where the effect of developmental stage proved to be insignificant. Lethal time values were estimated via probit analysis. Values of LT₅₀ ranged between 0.7 and 1.0 h for T. confusum, 0.7-1.9 h for O. surinamensis and 1.2-3.4 h for T. granarium. Our results are discussed with findings from relevant studies and the possibility of including extreme cooling to IPM programs against stored product pests is investigated.     

Insecticidal efficacy of phosphine fumigation at low pressure against major stored-product insect species in a commercial dried fig processing facility

We investigated the application of phosphine at low pressure for various exposure durations against major stored-product insects in a commercial dried fig processing facility in Central Greece. Trials were carried out inside a chamber, in which phosphine, in the form of aluminium phosphide pellets, was introduced with the use of a phosphine generator. The generator unit was also equipped with a vacuum pump to achieve low pressure inside the chamber. The chamber was filled with pallets with boxes containing figs. The insects tested were Tribolium confusum (all life stages), Ephestia elutella (eggs and larvae), Sitophilus oryzae (adults), Sitophilus granarius (adults), Rhyzopertha dominica (adults), Oryzaephilus surinamensis (adults) and Prostephanus truncatus (adults). Moreover, wheat grains containing immature stages of S. oryzae were also used. All insect-life stage combinations were exposed to phosphine at low pressure for 18, 48 and 72 h. In most cases, significant differences in mortality of insects treated with phosphine at low pressure compared to the control treatments were recorded. However, complete control (100%) was recorded only in the case of O. surinamensis adults and T. confusum larvae after exposure for 48 and 72 h, respectively. We conclude that the combined application of phosphine and low pressure at short exposure durations (up to 72 h) cannot provide sufficient control at least against the stored-product insect species and life stages tested in the present study.     

Comparison of spinetoram,imidacloprid, thiamethoxam and chlorantraniliprole against life stages of Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) on concrete

Spinetoram, imidacloprid, thiamethoxam and chlorantraniliprole are new insecticides with novel mode of actions, low mammalian toxicity and low impact to environment. In the present study, the efficacy of these insecticides was tested against Tribolium confusum Jacquelin du Val on concrete. Among the tested insecticides, spinetoram proved to be more effective, providing complete control of T. confusum adults and young larvae after 14 days of exposure. For the young larvae, thiamethoxam at the highest dose and chlorantraniliprole at both doses were equally effective with spinetoram. On the other hand, none of the tested insecticides were able to control T. confusum pupae. Moreover, none of the insecticides had ovicidal effect, with the exception of chlorantraniliprole in some combinations. From the mobile life stages, the most tolerant life stages were old larvae and the most susceptible young larvae. The presence of food (flour) moderated T. confusum mortality. From the results of the present study, we can conclude that spinetoram, thiamethoxam and chlorantraniliprole showed potential and need be further evaluated for surface treatments in stored product facilities. Our work underlined the need for good cleaning and sanitation procedures in warehouses and food processing facilities.     

Repellent effects of five monoterpenoid odours against Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.) in Calabar, Nigeria

Secondary plant compounds are recognised as important components of plant defence system against herbivores and pathogens. Five monoterpenoids, (R)-linalool, 1,8-cineole, (S)-2-heptyl acetate, (S)-2-heptanol and citral, which are natural components of the essential oils of Aframomum melegueta (K. Schum) and Zingiber officinale (Roscoe), were tested at the ratios in which they occur naturally for repellent activity against Tribolium castaneum (Herbst.) and Rhyzopertha dominica (F.) in a 4-way olfactometer. The results showed the repellent properties of the compounds as both beetles spent less time in the olfactometer arm containing the test stimuli. (R)-linalool and (S)-2-heptanol were stronger repellent compounds than the others. Linalool showed good repellent activity against T. castaneum (P = 0.001) as the insect spent 1.22 min in the test arm compared to the control arms (2.78 min), and R. dominica (P = 0.001) with 0.89 min in the test arm compared to 2.87 min in the control arms. With (S)-2-heptanol, T. castaneum spent 1.23 min in the test arm compared to 2.83 min in the control arms. R. dominica spent 1.61 min in the test arm and 2.69 min in the control arms. For the number of entries or visits made, while both insects were significantly repelled (P < 0.05) by the linalool-treated arm than the control, only R. dominica was repelled by the (S)-2-heptanol-treated arm (P = 0.038) compared to the control arms. The results indicate that A. melegueta and Z. officinale essential oils and their components could be suitable as safer repellents or fumigants against T. castaneum and R. dominica.     

Toxicity of essential oils isolated from three Artemisia species and some of their major components to granary weevil,Sitophilus granarius (L.) (Coleoptera: Curculionidae)

The essential oils of aerial parts of three Artemisia species (A. absinthium, A. santonicum and A. spicigera) were isolated by hydrodistillation method and tested for their toxicity against to granary weevil, Sitophilus granarius (L.) (Coleoptera: Curculionidae). All of the essential oils tested were found to be toxic to adults of S. granarius. The oils showed about 80–90% mortality of granary weevil, S. granarius at a dose of 9 μl/l air after 48 h of exposure. The constituents of these oils isolated from Artemisia spp. were also analyzed by GC–MS method. Camphor, 1,8-cineole, chamazulene, nuciferol propionate, nuciferol butanoate, caryophyllene oxide, terpinen-4-ol, borneol and α-terpineol were the predominant components of the oils. Some pure compounds (camphor, 1,8-cineole, terpinen-4-ol, borneol, bornyl acetate and α-terpineol) identified as major component in the oils, at 0.5, 0.75 and 1 μl/l air doses were also tested for their toxicity against S. granarius. While all pure compounds were found to be toxic against S. granarius, 1,8-cineole and terpinen-4-ol were more toxic among the tested pure compounds. 1,8-Cineole and terpinen-4-ol showed 100% mortality at all doses after 12 h of exposure. It can be concluded that essential oils of three Artemisia species and their major components, 1,8-cineole and terpinen-4-ol are potential control agents against S. granarius.     

Detection and characterization of bismerthiazol-resistance of Xanthomonas oryzae pv. oryzae

The resistance to bismerthiazol in the Xanthomonas oryzae pv. oryzae population in China has not been clearly determined. Three bismerthiazol-resistant mutants (1-1-1, 2-1-1, 4-1-1) of X. oryzae pv. oryzae were induced on rice plants and the characteristics were compared with their parental strain ZJ173. After 10 subcultures on fresh bismerthiazol-free nutrient agar plates and after being inoculation onto and re-isolation from untreated rice plants two times, the mutants remained pathogenicity on rice plants treated with 300 μg/ml bismerthiazol, indicating that the in vivo-induced bismerthiazol resistance was stable in vivo. The in vitro characteristics of the bismerthiazol-resistant mutants and a wild-type bismerthiazol-sensitive strain (ZJ173) were also compared. The three mutants grew faster than ZJ173 during the logarithmic growth phase but at the same rate during the decline phase. Bismerthiazol had protective and curative activity against strain ZJ173 but not against the three mutants. To evaluate the current degree of bismerthiazol resistance in field populations of X. oryzae pv. oryzae in China, 781 isolates of X. oryzae pv. oryzae were obtained from 1419 leaf samples collected from Yunnan, Sichuan, Jiangsu, Anhui, Hubei, Guangdong, Hainan, and Hunan Provinces in China from 2007 to 2009. The in vivo sensitivity of 505 of the 781 isolates to bismerthiazol was determined by applying them to rice plants treated with 300 μg/ml bismerthiazol and evaluating blast severity. The tested strains were considered resistant, if they showed <70% inhibition relative to strain ZJ173. Of the 505 isolates, 62 were resistant to bismerthiazol. The resistance frequencies among field population ranged from 0 to 21.1%, differed among the provinces, and was 11.2, 20.0 and 10.4% in 2007, 2008 and 2009, respectively. Resistance determined in vivo was not related to resistance determined in vitro. The in vivo-induced bismerthiazol resistance identified in this study will serve as a base line for further studying bismerthiazol resistance in fields, and the current degree of bismerthiazol resistance in field populations of X. oryzae pv. oryzae in China determined would be useful for the rice growers for control of rice bacterial blight.     

Effect of grain type on the insecticidal efficacy of SilicoSec against Sitophilus oryzae (L.) (Coleoptera: Curculionidae)

The insecticidal efficacy of the diatomaceous earth formulation SilicoSec^® (Agrinova GmbH, Germany) against the rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae) was evaluated on peeled rice, paddy rice, maize and barley in laboratory tests. SilicoSec was applied at five dose rates: 0.125, 0.25, 0.5, 1 and 1.5 g/kg of grain. The treated grains were infested with S. oryzae adults, and the weevil mortality was estimated after 24, 48 h, 7 and 14 d of exposure on treated grain. After the 14 d counts, all the exposed adults were removed and progeny production on treated grains was assessed 45 and 90 d later. Insecticidal efficacy of SilicoSec against the rice weevil was highly affected by grain type, exposure interval and dose rate. In all grains tested, longer exposure intervals increased weevil mortality. The efficacy of SilicoSec on maize was not satisfactory, given that after a 14 d exposure at the highest dose rate the mortality did not exceed 65%. Moreover, progeny production on treated maize was high, regardless of the dose rate. On the other hand, 100% of weevils were dead on barley treated with the two highest dose rates of SilicoSec, after a 7 d exposure. However, a complete suppression of progeny production on treated barley was not achieved, even at the highest dose rate. A satisfactory level of protection was recorded on paddy rice treated with 1 and 1.5 g of SilicoSec/kg. At these two rates, all adults were dead after a 7 d exposure, and no progeny were produced during the 90 d incubation period. In contrast, the mortality on peeled rice treated with 1.5 g of SilicoSec/kg did not reach 100%, even after 14 d of exposure, and a high number of progeny were recorded.     

Improvements in the qualities of gluten-free bread after using a protease obtained from Aspergillus oryzae

From a nutritional perspective, rice flour is one of the most valuable flours and it is suitable for preparing food for people suffering from wheat allergy. However, bread made from rice flour is very difficult to bake because it lacks gluten. We found that pre-fermenting of rice flour using Aspergillus oryzae facilitated a better formulation of gluten-free rice bread. Bread swelling was remarkably improved with a longer pre-fermenting period at 55 °C. The specific loaf volume (SLV) without polymeric thickeners after a 12 h fermentation was approximately 2.2-fold (2.98 ml/g) higher than that after 0 h (1.36 ml/g). An enzymatic assay of the batter indicated that protease activity during the pre-fermentation period increased from 0.38 to 1.44 U/ml and this activity correlated with bread swelling. Furthermore, a commercial protease from A. oryzae also produced similar results with an adequate SLV of 3.03 ml/g. Rheological analysis showed that batter treated with protease had an increased batter viscosity and decreased flour settling behavior because of the aggregation of flour particle after partial cleavage of storage proteins. These results indicated that the improved SLV was mainly because of an A. oryzae protease, which affected the batter rheology thereby improving gas retention before baking.     

Insecticidal effect of two novel pyrrole derivatives against two major stored product insect species

Several pyrrole compounds exhibit insecticidal properties against a wide range of insect pest species. In the present work, the insecticidal effect of the two new pyrrole derivatives, ethyl 3-(benzylthio)-4,6-dioxo-5-phenyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-carboxylate (3i) and isopropyl 3-(benzylthio)-4,6-dioxo-5-phenyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-carboxylate (3k) were studied as stored-grain protectants (maize and barley) against two major stored-product insect species, the confused flour beetle, Tribolium confusum Jaquelin du Val (Coleoptera: Tenebrionidae) adults and larvae and the Mediterranean flour moth, Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) larvae at three doses (0.1, 1 and 10 ppm) and five exposure intervals (1, 2, 7, 14 and 21d). All T. confusum adults were dead at all doses on barley treated with 3i after 21d of exposure, while for 3k mortality was >92%. Progeny production was very low (≤1 individual per vial) at all doses for both pyrrole derivatives. Mortality of T. confusum larvae was not complete with any dose of both pyrrole derivatives but it exceeded 96% with 3k at 10 ppm after 21d of exposure on maize. For barley, all exposed larvae were found dead at all doses of both pyrrole derivates after 7d of exposure. Mortality of E. kuehniella larvae was complete at 1 and 10 ppm of 3i and all doses of 3k at 7d of exposure. After 14d of exposure, all E. kuehniella larvae were dead at 0.1 ppm of 3i. The results of the present study indicate that the tested compounds have elevated insecticidal effect against both species tested on certain combinations of dose, exposure interval and type of commodity.     

Rheological and textural properties of tef [Eragrostis tef (Zucc.) Trotter] grain flour gels

Interest in tef [Eragrostis tef (Zucc.)Trotter] grain in food applications has increased in recent years because of its nutritional merits and the absence of gluten. With the objective of evaluating the suitability of tef for making gel type food products, gel viscoelastic properties of three varieties of tef (one brown and two white) at different concentrations (6, 8, 10, 12 & 14% w/w) were evaluated at 25 °C and 90 °C. The texture and color evolution for 16% (w/w) gels were evaluated. Proximate compositions of the flours were quantified. Rice, refined and whole wheat flours were analyzed as reference. The minimum flour concentration required for gel formation from the three tef varieties was 6–8%, similar to wheat flour. All tef flour suspensions pre-heated to 95 °C led to gels with a solid-like behavior (G′ > G″), both at 25 °C and 90 °C, with higher consistency than wheat gels at the same concentration. The dependence of viscoelastic moduli with concentration fulfilled the power law. The Avrami model was successfully fitted to the textural evolution of tef gels. Important differences were observed among tef and rice and wheat flours, probably contributed by their differences in protein, starch, lipid and fiber constituents. Gelling properties characterized suggest that tef flours would be suitable ingredients in gel food formulations.     

Low susceptibility of Spodoptera cosmioides,Spodoptera eridania and Spodoptera frugiperda (Lepidoptera: Noctuidae) to genetically-modified soybean expressing Cry1Ac protein

Spodoptera cosmioides (Walker), Spodoptera eridania (Stoll) and Spodoptera frugiperda (J. E. Smith) have caused significant damage on soybean Glycine max (L.) Merrill in Brazil. Genetically-modified MON 87701 × MON 89788 soybean that expresses the Cry1Ac protein is potentially an alternative tool for the management of these species. Purified protein bioassays were done to evaluate the susceptibility of S. cosmioides, S. eridania and S. frugiperda to Cry1Ac protein. The level of efficacy of the Bt soybean plants in controlling these species was measured through laboratory and greenhouse trials under high artificial insect infestations. The biology of these insects was evaluated over their development cycles to understand their life history when fed on Bt soybean. Purified Cry1Ac protein at the maximum concentration tested (100 μg Cry1Ac mL⁻¹ diet) resulted in low mortality of S. cosmioides and S. eridania (<13%) and intermediate mortality of S. frugiperda (50%). No significant effects of the Bt soybean plants were observed in the life table parameters of S. cosmioides and S. eridania. However, S. frugiperda fed on Bt soybean plants had a prolonged larval stage (by 5 days), reduced larvae viability, increased mean generation time (by 8 days) and reduced intrinsic rate of increase. In general, the Bt soybean plants showed poor control of Spodoptera species when evaluated by leaf-disc bioassay and greenhouse trials. Consequently, other control tactics must be used in combination with MON 87701 × MON 89788 soybean in the field for the efficient management of S. cosmioides, S. eridania and S. frugiperda.     

The early classification of the nutritional value of protein sources by biological tests using two insects found in stored foodstuffs of theTenebrionidae family:Tenebrio molitor L. andTribolium confusum Duval

Biological assays on white rats and chickens, used to screen the nutritional value of plants proteins, are compared with those of yellow mealworm (Tenebrio molitor L.) and confused flour beetle (Tribolium confusum Duval). With the twenty diets tested, the classification by vertebrates and insect larvae was largely similar.     

Screening of Uruguayan plants for deterrent activity against insects

We evaluated the anti-insectan activity of extracts from different vegetative parts of ten plant species native to Uruguay. The selected plants belong to five families: Bignoniaceae: Clytostoma callistegioides, Dolichandra cynanchoides, Macfadyena unguis-cati; Sapindaceae: Dodonaea viscosa, Allophylus edulis, Serjania meridionalis; Lamiaceae: Salvia procurrens, Salvia guaranitica; Solanaceae: Lycium cestroides; and Phytolaccaceae: Phytolacca dioica. The extracts were evaluated in independent bioassays against four insect pests and one beneficial insect. Aphid settling inhibition was evaluated with a grass specialist, Rhopalosiphum padi, and a feeding generalist, Myzus persicae (both Hemiptera: Aphididae). Antifeedant activity was tested with adults of the specialist Epilachna paenulata (Coleoptera: Coccinellidae) and larvae of the generalist Spodoptera littoralis (Lepidoptera: Noctuidae). Finally, contact toxicity was assessed with honey bees, Apis mellifera (Hymenoptera: Apidae). Strong settling inhibition (SI) activity (expressed as %SI, where 100% means complete inhibition by the extract) was found only for the twig extracts of A. edulis (Sapindaceae) against M. persicae (%SI = 77 ± 4). Antifeedant activity (expressed as % of feeding reduction (FR), where 100% means no consumption on extract-treated diet) against E. paenulata was significant for the leaf extracts of L. cestroides (Solanaceae) (%FR = 100 ± 0) as well as of all Bignoniaceae and Sapindaceae species. No extracts were active against S. littoralis larvae, and most of them were innocuous to honey bees, with the exception of L. cestroides and S. meridionalis leaf extracts.     

Weed control efficacy and winter wheat safety of a novel herbicide HW02

HW02, a pyruvate dehydrogenase inhibitor, is a newly developed herbicide for broadleaf weed control in wheat, maize and turf in China. Greenhouse and field experiments were conducted to evaluate its efficacy against weeds and safety to winter wheat. In the greenhouse experiment, this herbicide had higher activities than 2,4-D against Descurainia sophia (L.) Schur., Amaranthus retroflexus L., Capsella bursa-pastoris (L.) Medic., and Malachium aquaticum (L.) Fries.. When it was applied at late tillering stage of winter wheat in spring, the herbicide provided weed biomass reduction of 98%–100% at the rates 225–525 g a.i. ha⁻¹ and was safe to the crop at the rate of up to 900 g a.i. ha⁻¹. These results showed HW02 could be an alternative herbicide for resistant weed management because its mode of action is different from herbicides presently used.     

Effects of nitrogen on egg-laying inhibition and ovicidal response in planthopper-resistant rice varieties

A series of experiments was set up to examine the effects of nitrogen on rice (Oryza sativa L.) resistance against Nilaparvata lugens (Stål) and Sogatella furcifera (Horváth). Egg laying by N. lugens was reduced on the indica variety IR60. Nymph biomass (N. lugens and S. furcifera) was also lower on IR60: this was associated with low honeydew production and a high proportion of xylem-derived honeydew in N. lugens but not in S. furcifera. Nitrogen increased egg-laying by S. furcifera and increased N. lugens nymph biomass on all varieties tested. Oviposition and egg mortality in both planthopper species were examined on plants at 15, 30 and 45 days after sowing (DAS). Sogatella furcifera laid more eggs on plants at 15 DAS, but laid few eggs during darkness; N. lugens continued to lay eggs on older rice plants (30 DAS) and during darkness. Egg mortality was high on cv. Asiminori, highest at 45 DAS, and higher for S. furcifera than for N. lugens. Mortality of S. furcifera eggs was associated with lesions around the egg clusters. These were more common around clusters laid during the day and suggested induction by Asiminori of an ovicidal response. Egg mortality declined under higher soil nitrogen levels. Results are discussed in the light of improving rice resistance against planthoppers and reducing rates of planthopper adaptation to resistance genes.     

Proteomic analysis of the mature kernel aleurone layer in common and durum wheat

The aleurone layer (AL) is one of inner tissues removed from the grain with the wheat bran. It is the main source of vitamins, minerals and antioxidants of potential nutritional value in the wheat kernel. The AL of three varieties of each of the two main species of wheat, Triticum aestivum (ABD) and Triticum durum (AB), were manually dissected and analysed using two-dimensional gel-based proteomics. A total of 1258 and 1109 Coomassie-stained spots were detected in the AL of representatives of the ABD and AB genomes. In two varieties (T. aestivum Chinese Spring and T. durum Bidi17), grown in two different years with full fungicide protection, no quantitative or qualitative (presence/absence) differences in spots were detected, suggesting that AL proteome is strongly genetically controlled. Comparison within and between species revealed a total of 339 AL significant protein spots. Among these spots, 30.8% differed within T. aestivum and 56.5% within T. durum varieties, whereas only 12.7% differed between the two species. Among the 142 AL proteins identified using MALDI-TOF and LC-MS/MS, 57% were globulin type storage proteins (Glo-3, Glo-3B, Glo-3C, Glo-2), 16.2% were involved in carbohydrate metabolism and 17.6% in defence/stress pathways. These variations in AL proteome are discussed.     

Isolation of entomopathogenic Bacillus from a biodynamic olive farm and their pathogenicity to lepidopteran and coleopteran insect pests

The occurrence of Bacillus entomopathogenic bacteria on a Tunisian biodynamic farm was determined by examining 75 samples from olive tree (Olea europaea L.) habitats. A total of 40 Bacillus isolates were characterized according to their phenotypic, physiological and biochemical parameters. Isolates of the species Bacillus subtilis, Bacillus mycoides, Brevibacillus brevis, Paenibacillus polymyxa, Bacillus licheniformis, Bacillus sp. (1), Bacillus sp. (2) and a standard strain Btk HD-1 were used separately in feeding bioassays on fresh artificial diet against larvae of lepidopterans Prays oleae (Bernard) and Palpita unionalis (Hübner) and coleopterans Hylesinus oleiperda (F.) and Phloeotribus scarabaeoides (Bernard), which are olive tree pests. Larvae were successfully reared on an artificial diet with 25 g powdered olive tree leaves. Compared to the control data, only Btk and the isolates of B. licheniformis, P. polymyxa and B. brevis were entomopathogenic. Larval mortality assessed 7 days post-treatment showed high mortality rates with Btk to lepidopteran larvae (86.6% for P. oleae and 80.9% for P. unionalis) and low mortality against coleopteran pests. B. brevis isolates showed high mortality rates against P. oleae (up to 67.9%). B. licheniformis isolates caused up to 59.2% larval mortality for P. oleae and 43.6% for P. unionalis. Highest coleopteran mortality was achieved by P. polymyxa isolates (up to 55%). According to the 16S rDNA results, isolates of each of the three entomopathogenic strains were similar. Proteins in the strain supernatants were toxic to P. oleae larvae with LC50 values of 10.0 (B. brevis), 12.5 (B. licheniformis) and 37.6 μg/ml (P. polymyxa). Also, P. polymyxa showed an LC50 of 12.4 mg/l against P. scarabaeoides. Our results suggest that entomopathogenic Bacillus present locally in the biodynamic farm could be used in biological control programmes of olive tree pests.