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Comparative susceptibility of European corn borer, southwestern corn borer, and sugarcane borer (Lepidoptera: Crambidae) to Cry1Ab protein in a commercial Bacillus thuringiensis corn hybrid.

One field strain each of the European corn borer, Ostrinia nubilalis (Hübner); southwestern corn borer, Diatraea grandiosella Dyar; and sugarcane borer, Diatraea saccharalis (F.); were collected from cornfields in northeastern Louisiana. Susceptibilities of the field strain and a corresponding laboratory strain of the three borer species to Cry1Ab protein in DK69-70 Bacillus thuringiensis (Bt) corn hybrid were determined by exposing neonates to intact leaf tissues from whorl stage plants or by feeding neonates or third instars on a meridic diet treated with different concentrations of Cry1lAb protein extracted from Bt corn leaves. Mortality and growth of larvae were evaluated after 2 and 4 d posttreatment in the bioassays by using intact leaf tissues or after 7 d in the bioassays by using diet incorporating Cry1Ab protein. D. saccharalis was the least susceptible species to Cry1Ab protein among the three species, followed by D. grandiosella, whereas O. nubilalis was most susceptible. The 2-d mortality of D. saccharalis neonates on intact Bt leaf tissues was lower than that of O. nubilalis and D. grandiosella. All neonates of O. nubilalis were killed on the diet treated with Cry1Ab protein at 0.5 and 1 mg/kg. The mortality of D. grandiosella was > 75% at 1 mg/kg, but it was < 6% for D. saccharalis at 1 mg/kg. The LC50 values of D. saccharalis were 3- and 11-fold higher than those of D. grandiosella and O. nubilalis, respectively. The LC90 values of D. saccharalis were 8- and 32-fold higher than those of D. grandiosella and O. nubilalis, respectively. Larval growth of the three species on Cry1Ab-treated diet was inhibited, but the inhibition was greater for O. nubilalis and D. grandiosella than for D. saccharalis. The lower susceptibility of D. saccharalis to Cry1Ab protein suggests that it is necessary to verify if a high-dose Bt corn for O. nubilalis and D. grandiosella is also a high dose for D. saccharalis.

Analysis of Variance↗

Bioinformatics correctly identifies many type III secretion substrates in the plant pathogen Pseudomonas syringae and the biocontrol isolate P. fluorescens SBW25.

The plant pathogen Pseudomonas syringae causes disease by secreting a potentially large set of virulence proteins called effectors directly into host cells, their environment, or both, using a type III secretion system (T3SS). Most P. syringae effectors have a common upstream element called the hrp box, and their N-terminal regions have amino acids biases, features that permit their bioinformatic prediction. One of the most prominent biases is a positive serine bias. We previously used the truncated AvrRpt2(81-255) effector containing a serine-rich stretch from amino acids 81 to 100 as a T3SS reporter. Region 81 to 100 of this reporter does not contribute to the secretion or translocation of AvrRpt2 or to putative effector protein chimeras. Rather, the serine-rich region from the N-terminus of AvrRpt2 is important for protein accumulation in bacteria. Most of the N-terminal region (amino acids 15 to 100) is not essential for secretion in culture or delivery to plants. However, portions of this sequence may increase the efficiency of AvrRpt2 secretion, delivery to plants, or both. Two effectors previously identified with the AvrRpt2(81-255) reporter were secreted in culture independently of AvrRpt2, validating the use of the C terminus of AvrRpt2 as a T3SS reporter. Finally, using the reduced AvrRpt2(101-255) reporter, we confirmed seven predicted effectors from P. syringae pv. tomato DC3000, four from P. syringae pv. syringae B728a, and two from P. fluorescens SBW25.

Amino Acid Sequence↗

Sex pheromones and reproductive isolation of the lesser peachtree borer and the peachtree borer.

(E,Z)-3,13-octadecadien-1-ol acetate, and (Z,Z)-3,13-octadecadien-1-ol acetate, isolated from the female lesser peachtree borer, Synanthedon pictipes (Grote and Robinson), and the female peachtree borer, Sanninoidea exitiosa (Say), respectively, strongly attract the respective males of these species in field bioassays. These compounds are the largest pheromones isolated thus far from a lepidopterous species. Sanninoidea exitiosa males did not respond to the synthesized (E,Z)-isomer, and low concentrations of it in the synthesized (Z,Z)-isomer did not interfere with their response to the (Z,Z)-isomer. In contrast, even very low concentrations of the (Z,Z)-isomer (1 percent) in the (E,Z)-isomer significantly inhibited the response of Synanthedon pictipes males.

Animals↗

Nontarget effects--the Achilles' heel of biological control? Retrospective analyses to reduce risk associated with biocontrol introductions.

Controversy exists over ecological risks in classical biological control. We reviewed 10 projects with quantitative data on nontarget effects. Ten patterns emerged: (a) Relatives of the pest are most likely to be attacked; (b) host-specificity testing defines physiological host range, but not ecological range; (c) prediction of ecological consequences requires population data; (d) level of impact varied, often in relation to environmental conditions; (e) information on magnitude of nontarget impact is sparse; (f) attack on rare native species can accelerate their decline; (g) nontarget effects can be indirect; (h) agents disperse from agroecosystems; (i) whole assemblages of species can be perturbed; and (j) no evidence on adaptation is available in these cases. The review leads to six recommendations: Avoid using generalists or adventive species; expand host-specificity testing; incorporate more ecological information; consider ecological risk in target selection; prioritize agents; and pursue genetic data on adaptation. We conclude that retrospective analyses suggest clear ways to further increase future safety of biocontrol.

Animals↗

Large-scale management of insect resistance to transgenic cotton in Arizona: can transgenic insecticidal crops be sustained?

A major challenge for agriculture is management of insect resistance to toxins from Bacillus thuringiensis (Bt) produced by transgenic crops. Here we describe how a large-scale program is being developed in Arizona for management of resistance to Bt cotton in the pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), and other insect pests of cotton. Financial support from growers makes this program possible. Collaboration between the Arizona Cotton Research and Protection Council, the University of Arizona, and government agencies has led to development of resistance management guidelines, a remedial action plan, and tools for monitoring compliance with the proposed guidelines. Direct participation in development of resistance management policies is a strong incentive for growers to invest in resistance management research. However, more research, regularly updated regulations, and increased collaboration between stakeholders are urgently needed to maintain efficacy of Bt toxins in transgenic crops.

Animals↗

Potential of Agrotis ipsilon nucleopolyhedrovirus for suppression of the black cutworm (Lepidoptera: Noctuidae) and effect of an optical brightener on virus efficacy.

Studies were performed in the laboratory, greenhouse and field to assess the potential of Agrotis ipsilon multicapsid nucleopolyhedrovirus (AgipMNPV) and a viral enhancing agent, M2R, for suppression of Agrotis ipsilon (Hufnagel). In laboratory droplet feeding bioassays, AgipMNPV was shown to be highly active against third-instar A. ipsilon. The optical brightener M2R significantly reduced LD50 estimates by approximately 160-fold, but had no direct effect on survival time estimates. In greenhouse trials, spray and bait formulations of AgipMNPV significantly reduced feeding damage to corn seedlings caused by third-instar A. ipsilon. In two sets of replicated field trials, bait formulations of AgipMNPV significantly reduced feeding damage to corn seedlings by third-instar A. ipsilon. However, there were no beneficial effects attributable to the inclusion of M2R in AgipMNPV formulations under greenhouse or field conditions. It seems likely that in an appropriately designed pest management program AgipMNPV could be used to suppress field populations of early and mid-instar A. ipsilon.

Animals↗

Evaluation of granular corncob formulations of Bacillus thuringiensis serovar israelensis against mosquito larvae using a semi-field bioassay method.

A semi-field bioassay method using commonly available laundry tubs evaluated efficacy and residual activity of 6 granular corncob formulations of Bacillus thuringiensis serovar israelensis against laboratory-reared late 3rd- and early 4th-instar larvae of Aedes taeniorhynchus introduced to the tubs. These formulations produced appreciable larval mortalities (up to 75%) for 9 days posttreatment, with indications of having more prolonged activity. The consistent posttreatment larval mortality trends and the elucidated efficacy differences between formulations suggest that this bioassay method for the granular formulations is reliable and reproducible.

Aedes↗

Inhibition effect of transgenic tobacco plants expressing snowdrop lectin on the population development of Myzus persicae.

The cDNAs of snowdrop lectin mature protein and its precursor protein, GNA12 and GNA34, were inserted downstream of a 35S promoter with a double enhancer and a "omega" fragment of TMV-RNA cDNA in the binary vector pBin438, or the phloem-specific CoYMV promoter in the vector pBcop1, respectively, resulting in the construction of four plant expression vectors pBGna12, pBGna34, pBCGna12, and pBCGna34. Leaf stripes of Nicotiana tabacum var. K326 were transformed with A. tumefaciens LBA4404 harboring the above expression vectors, respectively. PCR and Southern blot analysis showed that foreign GNA genes were inserted into the genome of transformed tobacco plants. Western blot analysis indicated that GNA could be expressed efficiently up to 0.08-0.15% of total soluble proteins in transgenic tobacco plants with pBGna34 or pBCGna34, while in those with pBGna12 and pBCGna12 GNA were hardly detected by immunoassay. The results from insect bioassay with a peach aphid (Myzus persicae) demonstrated that the transgenic plants of pBGna34 and pBCGna34 were aphid-resistant as shown by a 45-60% reduction in insect population density, with some individual transgenic lines being reduced by over 90%. In addition, it was evident that the 35S promoter with a double enhancer and CoYMV promoter had similar abilities to direct the GNA gene to express in transgenic tobacco plants, but because the CoYMV promoter drives the foreign gene in a phloem-specific expression manner, the transgenic plants of pBCGna34 showed higher aphid resistance.

Animals↗

Glandular trichome extracts from Medicago sativa deter settling by the potato leafhopper Empoasca fabae.

Extracts of glandular trichomes from Medicago sativa were tested for their ability to disrupt the settling behavior of the potato leafhopper, Empoasca fabae. Erect and procumbent glandular trichomes were mechanically isolated from stem sections of resistant genotype "G98A," and nonglandular trichomes were collected from susceptible cultivar "Ranger." Isolated trichomes were extracted with chloroform, acetone, and ethanol, and the resulting crude extracts were applied to the surface of a sachet containing an artificial diet. Leafhoppers were offered a two-way choice between crude trichome extracts from G98A and Ranger. All three of G98A solvent extracts caused various degrees of diet rejection, resulting in the crude Ranger trichome extracts being preferred over G98A extracts. Overall, the fewest leafhoppers settled on the ethanolic extracts. Additional bioassays documented a dose response associated with G98A ethanolic extracts when compared with Ranger trichome extracts and a solvent control. No difference in preference behavior was detected between Ranger trichome extracts and a solvent control. Gas chromatography-mass spectrometry analysis revealed a homologous series of nonvolatile fatty acid amides C(n)H(2n+1)NO (n = 19-23) unique to G98A glandular trichome extracts.

Amides↗

Expression of the cry9Aa2 B.t. gene in tobacco chloroplasts confers resistance to potato tuber moth.

We report here the control of potato tuber moth (Phthorimaea operculella) by incorporating a truncated Bacillus thuringiensis cry9Aa2 gene in the plastid genome. Plasmids pSKC84 and pSKC85 are derivatives of a new polycistronic plastid transformation vector, pPRV312L, that carries spectinomycin resistance (aadA) as a selective marker and targets insertions in the trnI-trnA intergenic region. The Cry9Aa2 N-terminal region (82.1 kDa; 734 amino acids) was expressed in a cassette, which consists of 49 nucleotides of the cry9Aa2 leader and the 3'-untranslated region of the plastid rbcL gene (TrbcL), and relies on readthrough transcription from the plastid rRNA operon. In a tobacco leaf bioassay, expression of Cry9Aa2 conferred resistance to potato tuber moth. In accordance, the Cry9Aa2 insecticidal protein accumulated to high levels, approximately 10% of the total soluble cellular protein and approximately 20% in the membrane fraction. However, high-level Cry9Aa2 expression significantly delayed plant development. Thus, a practical system to control potato tuber moth by Cry9Aa2 expression calls for down-regulation of its expression.

Animals↗

[Initiation of auto-oscillations in a distributed system with multiple equilibria].

Possible alterations in time of population numbers due to abrupt nonuniformity of environmental conditions were shown by a mathematical model. Elimination of borders between the local systems with stable uniform regimes results in the appearance of auto-oscillations of large amplitude in the whole region. The effect is interpreted in terms of the consequences of applying chemicals for pest control, and of chemotherapy influence on the immune system as well.

Animals↗

Cyanide production by rhizobacteria and potential for suppression of weed seedling growth.

Rhizobacteria strains were characterized for ability to synthesize hydrogen cyanide and for effects on seedling root growth of various plants. Approximately 32% of bacteria from a collection of over 2000 isolates were cyanogenic, evolving HCN from trace concentrations to > 30 nmoles/mg cellular protein. Cyanogenesis was predominantly associated with pseudomonads and was enhanced when glycine was provided in the culture medium. Concentrations of HCN produced by rhizobacteria were similar to exogenous concentrations inhibiting seedling growth in bioassays, suggesting that cyanogenesis by rhizobacteria in the rhizosphere can adversely affect plant growth. Growth inhibition of lettuce and barnyardgrass by volatile metabolites of the cyanogenic rhizobacteria confirmed that HCN was the major inhibitory compound produced. Our results suggest that HCN produced in the rhizospheres of seedlings by selected rhizobacteria is a potential and environmentally compatible mechanism for biological control of weeds.

Culture Media↗

Sex ratio, emergence and infection rates of Heterotylenchus-exposed Musca autumnalis (Diptera: Muscidae).

Experimental infections were produced with the host-specific nematode Heterotylenchus autumnalis, which sterilizes its host, the face fly (Musca autumnalis). The high infection rate achieved (means = 82.5%) was similar for male and female flies. The pupal emergence rate was significantly lower for fly larvae exposed to the nematodes than for unexposed controls. Heterotylenchus-exposed flies had a sex ratio skewed in favor of males. This indicates that in addition to sterilization, H. autumnalis has other deleterious effects on its host. Heterotylenchus autumnalis is therefore a candidate for use as a potential biological control agent of the face fly.

Animals↗

Effect of stabilizers on the shelf-life of Penicillium frequentans conidia and their efficacy as a biological agent against peach brown rot.

Stabilizers were added to conidia of Penicillium frequentans at two different points of the production-formulation process to improve shelf-life of conidia stored at different temperatures. Effects were also tested on conidial germination and production. Germination of conidia without additives was 90.2%; sodium chloride, potassium chloride, triton TX100, dimethyl sulphoxide, peroxidase, 0.375% and 0.075% ascorbic acid, 7.5% and 3.75% sucrose, and 7.5% and 3.75% d-sorbitol reduced significantly (P=0.05) conidial germination, while no effect was observed with glucose, lactose, maltose, sodium glutamate, glycerol, peptone, sodium alginate, carboximethylcellulose, Tween 80, and gelatine. Production of P. frequentans conidia in solid-state fermentation without additives was 1.07 conidia x 10(8) g(-1) of dry substrate. The highest tested doses of glucose, lactose, maltose, sodium glutamate, and glycerol enhanced production of P. frequentans, while the lowest tested doses of d-sorbitol and ascorbic acid reduced it. No significant effect was observed with sucrose, peptone, sodium alginate, carboximethylcellulose, gelatine and Tween 80. Conidial germinability after one year of storage at different temperatures was studied in some formulations. It was lower than 18% after 365 days of storage at room temperature in control samples (without any additive), being enhanced when 7.5% glucose, 7.5% glycerol, or 1.5% sodium alginate was added to the substrate in bags before fermentation; or when 7.5% glucose, 7.5% sodium glutamate, or 1.5% sodium alginate was added to conidia before drying. Germinability of conidia produced without any additive and stored at 4 degrees C was significantly higher (38%) than at room temperature, being enhanced when 7.5% glycerol or 1.5% sodium alginate was added to the substrate in bags before fermentation; and when 7.5% glucose, or 1.5% sodium alginate was added to conidia before drying. No effect was observed with the presence or absence of light or high vacuum. Four formulations of P. frequentans conidia reduced disease incidence by more than 55%. The relationship of the disease control with the viability of P. frequentans was discussed.

Biological Factors↗

Comparative toxicity of three ecdysone agonist insecticides against the Mediterranean flour moth.

The Mediterranean flour moth, Ephestia Kuehniella Zeller (Lepidoptera: Pyralidae), is an important pest in stored products worldwide, and is one of the major pests in flour mills in Algeria. Because environmental consideration, alternative approaches to neurotoxic insecticides, as well as safe, effective, and sound integrated pest management strategies are developed pest control agents such as the insect growth regulator (IGRs). Among these IGRs, the bisacylhydrazine derivatives are nonsteroidal ecdysterold agonists that mimic the action of moulting hormones and induce a precocious and incomplete moult in several insect orders. In topical bioassays using the pupae of E. kuehniella, three ecdysteroid agonists: RH-5849, the first bisaclhydrazine ecdysone agonist and two analogs, RH-5992 (tebufenozide) and RH-0345 (halofenozide), were evaluated on the reproduction under laboratory conditions. In a first series of experiments, the efficacy of these compounds was tested. These compounds exhibited insecticidal activity and the duration of pupal development was reduced with a dose-response relationship. Among the three tested compounds, tebufenozide (LD50 = 0.005 microg) appeared the most potent ecdysteroid agonist against E. kuehniella (RH-5849: LD50 = 0.05 microg and RH-0345: LD50 = 5.10 microg). In a second series of experiments, the effects of the ecdysone agonists (LD50) were investigated on the reproduction. Data showed that the three compounds affected growth of ovaries as evidenced by morphometric measurements of the ovaries from newly emerged adult females. In addition, the thickness of the chorion from basal oocytes was reduced only by RH-5992 and RH-0345. However, electron microscopic observations revealed that the three compounds had no significant effect on the fine structure of chorion. Finally, measurements of ovarian ecdysteroids' production by an enzyme immunoassay showed an increase in the hormonal amounts recorded in treated series compared to control series.

Animals↗

Beauveria bassiana as a pathogen of the Mexican fruit fly (Diptera: Tephritidae) under laboratory conditions.

Bioassays were carried out under controlled conditions (27 +/- 2 degrees C, 80 +/- 5% RH, and a photoperiod of 12:12 [L:D] h) to evaluate the effect of eight strains of the entomopathogenic fungus Beauveria bassiana upon larvae, pupae, and adult females of the Mexican fruit fly, Anastrepha ludens (Loew). Mortality of the immature stages was low, 2-8% in larvae and 0% in pupae. However, very high levels of mortality were obtained for adult flies, with values of 100, 98, and 98% for the strains Bb16, Bb24, and Bb26, respectively. LC50 values for these three strains ranged from 3.12 x 10(6) to 9.07 x 10(6) conidia/ml. Lethal time 50 (LT50) was 2.8, 3.7, and 4.2 d for Bb16, Bb26, and Bb24 strains, respectively, with an average LT50 of 4.4 d across all strains. The fungal mycelium emerged through the soft parts of the exoskeleton, such as the wing bases, mouth, intersegmental regions of the legs, and membranous regions of the abdomen, coxae, and neck. Maximum percentage sporulation ranged from 66.4 to 74.7% for the three most virulent strains.

Animals↗