Conservation biology: Parasites lost.
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The rhizobacterium Pseudomonas chlororaphis PCL1391 produces the antifungal metabolite phenazine-1-carboxamide (PCN), which is a crucial trait in its competition with the phytopathogenic fungus Fusarium oxysporum f. sp. radicis-lycopersici in the rhizosphere. The expression of the PCN biosynthetic gene cluster in PCL1391 is population density-dependent and is regulated by the quorum-sensing genes phzI and phzR via synthesis of the autoinducer N-hexanoyl-L-homoserine lactone (C6-HSL). Here, we describe the identification of an additional regulatory gene of PCN biosynthesis in PCL1391. A mutation in the psrA gene (Pseudomonas sigma regulator), the gene product of which is a member of the TetR/AcrR family of transcriptional regulators, resulted in increased production of autoinducer molecules and PCN. Expression studies showed that inactivation of psrA resulted in increased expression of the phzI and phzR genes and the phz biosynthetic operon and that introduction of functional copies of psrA represses the expression of these genes, resulting in reduced production of autoinducer signal and PCN. Surprisingly, inactivation of psrA in the phzI or phzR quorum-sensing mutants, which do not produce detectable amounts of PCN and autoinducers by themselves, restored PCN biosynthesis. This phenomenon was accompanied by the appearance of compounds with autoinducer activities migrating at the positions of C4-HSL and C6-HSL on C18 reverse phase-thin-layer chromatography. These observations indicate that PsrA also represses at least one silent, yet unidentified, quorum-sensing system or autoinducer biosynthetic pathway in PCL1391. The expression of psrA declines at the onset of the stationary phase at the same moment at which quorum-sensing (-regulated) genes are activated. In addition, expression studies in a psrA- and a multicopy psrA background showed that psrA is autoregulated. Multiple copies of psrA repress its own expression. Mutation of gacS, encoding the sensor kinase member of a two-component global regulatory system significantly reduced production of autoinducers and PCN. We show a novel link between global regulation and quorum sensing via the PsrA regulator.
The refuge strategy is used widely for delaying evolution of insect resistance to transgenic crops that produce Bacillus thuringiensis (Bt) toxins. Farmers grow refuges of host plants that do not produce Bt toxins to promote survival of susceptible pests. Many modelling studies predict that refuges will delay resistance longest if alleles conferring resistance are rare, most resistant adults mate with susceptible adults, and Bt plants have sufficiently high toxin concentration to kill heterozygous progeny from such matings. In contrast, based on their model of the cotton pest Heliothis virescens, Vacher et al. (Journal of Evolutionary Biology, 16, 2003, 378) concluded that low rather than high toxin doses would delay resistance most effectively. We demonstrate here that their conclusion arises from invalid assumptions about larval concentration-mortality responses and dominance of resistance. Incorporation of bioassay data from H. virescens and another key cotton pest (Pectinophora gossypiella) into a population genetic model shows that toxin concentrations high enough to kill all or nearly all heterozygotes should delay resistance longer than lower concentrations.
Exploring the possibility of enhancing the properties of baculoviruses as biological control agents of insect pests, we tested the effect of expressing an insect gene (jhe) encoding juvenile hormone esterase. Juvenile hormone esterase inactivates juvenile hormone, which regulates the outcome of an insect molt. A cDNA encoding the juvenile hormone esterase of Heliothis virescens was inserted into the genome of Autographa californica nuclear polyhedrosis virus such that the gene was expressed under the control of a strong, modified viral promoter. This virus, however, naturally encodes an ecdysteroid UDP-glucosyltransferase which inactivates ecdysone, the hormone which initiates molting. Since ecdysteroid UDP-glucosyltransferase could mask the effects of jhe expression by blocking molting entirely, jhe-expressing viruses in which the ecdysteroid UDP-glucosyltransferase gene was deleted or disrupted were constructed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of proteins from infected cells revealed several intracellular proteins and two major secreted proteins which reacted with antibodies to authentic juvenile hormone esterase. Western blot analysis coupled with tunicamycin treatment indicated that differential glycosylation was responsible for the multiple products. Hemolymph of recombinant virus-infected fourth-instar Trichoplusia ni larvae contained levels of juvenile hormone esterase activity 40-fold higher than maximal levels found in uninfected larvae. However, little or no difference in developmental characteristics, weight gain, or time of mortality was observed between insects infected with the jhe-expressing viruses and control viruses.
Insecticidal crystal proteins from Bacillus thuringiensis in sprays and transgenic crops are extremely useful for environmentally sound pest management, but their long-term efficacy is threatened by evolution of resistance by target pests. The diamondback moth (Plutella xylostella) is the first insect to evolve resistance to B. thuringiensis in open-field populations. The only known mechanism of resistance to B. thuringiensis in the diamondback moth is reduced binding of toxin to midgut binding sites. In the present work we analyzed competitive binding of B. thuringiensis toxins Cry1Aa, Cry1Ab, Cry1Ac, and Cry1F to brush border membrane vesicles from larval midguts in a susceptible strain and in resistant strains from the Philippines, Hawaii, and Pennsylvania. Based on the results, we propose a model for binding of B. thuringiensis crystal proteins in susceptible larvae with two binding sites for Cry1Aa, one of which is shared with Cry1Ab, Cry1Ac, and Cry1F. Our results show that the common binding site is altered in each of the three resistant strains. In the strain from the Philippines, the alteration reduced binding of Cry1Ab but did not affect binding of the other crystal proteins. In the resistant strains from Hawaii and Pennsylvania, the alteration affected binding of Cry1Aa, Cry1Ab, Cry1Ac, and Cry1F. Previously reported evidence that a single mutation can confer resistance to Cry1Ab, Cry1Ac, and Cry1F corresponds to expectations based on the binding model. However, the following two other observations do not: the mutation in the Philippines strain affected binding of only Cry1Ab, and one mutation was sufficient for resistance to Cry1Aa. The imperfect correspondence between the model and observations suggests that reduced binding is not the only mechanism of resistance in the diamondback moth and that some, but not all, patterns of resistance and cross-resistance can be predicted correctly from the results of competitive binding analyses of susceptible strains.
We constructed a model for Bacillus thuringiensis Cry1 toxin binding to midgut membrane vesicles from Heliothis virescens. Brush border membrane vesicle binding assays were performed with five Cry1 toxins that share homologies in domain II loops. Cry1Ab, Cry1Ac, Cry1Ja, and Cry1Fa competed with (125)I-Cry1Aa, evidence that each toxin binds to the Cry1Aa binding site in H. virescens. Cry1Ac competed with high affinity (competition constant [K(com)] = 1.1 nM) for (125)I-Cry1Ab binding sites. Cry1Aa, Cry1Fa, and Cry1Ja also competed for (125)I-Cry1Ab binding sites, though the K(com) values ranged from 179 to 304 nM. Cry1Ab competed for (125)I-Cry1Ac binding sites (K(com) = 73.6 nM) with higher affinity than Cry1Aa, Cry1Fa, or Cry1Ja. Neither Cry1Ea nor Cry2Aa competed with any of the (125)I-Cry1A toxins. Ligand blots prepared from membrane vesicles were probed with Cry1 toxins to expand the model of Cry1 receptors in H. virescens. Three Cry1A toxins, Cry1Fa, and Cry1Ja recognized 170- and 110-kDa proteins that are probably aminopeptidases. Cry1Ab and Cry1Ac, and to some extent Cry1Fa, also recognized a 130-kDa molecule. Our vesicle binding and ligand blotting results support a determinant role for domain II loops in Cry toxin specificity for H. virescens. The shared binding properties for these Cry1 toxins correlate with observed cross-resistance in H. virescens.
Aphidophagous predators such as hoverfly and ladybird beetles are effective biological agents to control aphid pests in perennial and annual cultivated species. Introduction and conservation of beneficial insects are two ways to increase natural control of pests. Whether massive releases of entomophagous insects are expensive and time consuming, the preservation of predator natural populations can be expected by reducing and by adapting chemical treatments in crop fields. Vegetable cultivated areas increased in Belgium for several years, mainly Fabaceae species such as peas and beans. In this work, the evolution of ladybird species population was assessed from May to June in broad bean fields (Vicia faba L.) between Waremme and Hannut, in Hesbaye. Weekly, the aphid and aphidophagous beetle populations were collected from yellow traps and determined on plants by visual observations. Even if five ladybird species were identified, three of them represented more than 95% of the collected insects (Coccinella septempunctata L., Propylea quatuor-decimpunctata L. and Psyllobora vingintiduopunctata L.). Evolution of coccinellid populations during the cultivation season was discussed in relation to the presence of potential aphid preys and the agrochemical treatments which were applied. Integrated pest management in vegetable fields constitute a reliable way to increase the quality level of fresh vegetables in terms of pesticide residue limitations.
The study on the nuclear polyhedral virus epidemic of Spodoptera litura under effects of temperature showed that at above 29 degrees C, S. litura larvae started to die 2-3 d after virus-inoculation, and reached the peak 4-5 d after the inoculation. The mortality and death velocity of virus-infected host larvae were increased with increasing incubation temperature. No thermo-inhibition temperature for the virus was observed in the test temperature range, while there existed a constant temperature range, i.e., 29-35 degrees C, for the highest mortality of infected host larvae. Complementary log-log regression model could well simulate the daily distribution of host larvae mortality, and the cumulative disease death time could be described by Gompertz model. The revised biophysical model could well simulate the relationship between S. litura larvae mortality and temperature, and be applied to virus epidemic forecasting.
A possible method for genetic control of insect vector species involves the use of translocation heterozygotes. The potential of single and double heterozygotes already available in Aedes aegypti has been investigated with computer simulations of release strategies. Such simulations indicate a possible role for translocation heterozygotes of these types in an insect population characterized by a 5-fold population growth per generation, or less.
As growers switch to transgenic crops and selective insecticides that are less toxic to natural enemies, natural enemies can become more important in agricultural pest management. Current decision-making guides are generally based on pest abundance and do not address pest and natural enemy toxicity differences among insecticides or the impact of natural enemies on pest survival. A refined approach to making pest management decisions is to include the impact of natural enemies and insecticides, thereby better integrating biological and chemical control. The result of this integration is a dynamic threshold that varies for each product and the level of biological control expected. To demonstrate the significance of conserved biological control in commercial production, a decision-making guide was developed that evaluates control options for European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), in sweet corn, Zea mays L., where the primary natural enemies are generalist predators. Management options are lambda-cyhalothrin (broad-spectrum insecticide), spinosad (selective insecticide), Trichogramma ostriniae (Peng and Chen) (Hymenoptera: Trichogrammatidae) (parasitoid), and Bacillus thuringiensis (Bt) sweet corn (transgenic variety). The key factors influencing thresholds for all treatments are the intended market, predator populations, and the presence of alternative foods for the predators. Treatment cost is the primary factor separating the threshold for each treatment within a common scenario, with the lowest cost treatment having the lowest pest threshold. However, when the impact of a treatment on natural enemies is projected over the 3-wk control period, the impact of the treatment on predators becomes the key factor in determining the threshold, so the lowest thresholds are for broad-spectrum treatments, whereas selective products can have thresholds > 6 times higher by the third week. This decision guide can serve as a framework to help focus future integrated pest management research and to aid in the selection of pest management tools.
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Data from nine trials conducted from 1990 to 1998 in apple orchards in Nova Scotia and Quebec, Canada, were used to estimate the predator-prey selectivity of miticides and their potential compatibility with biological control of mites. The European red mite Panonychus ulmi (Koch) was the dominant and more harmful phytophagous species, followed by the apple rust mite, Aculus schlechtendali (Nalepa). Two predacious mites, the phytoseiid, Typhlodromus pyri Sheuten, and the stigmaeid, Zetzellia mali (Ewing), were often found in the orchards. We used one minus the ratio of mite-days in treated plots to those in the control plots as an index of population suppression and toxicity of the miticides. Miticides were then categorized into classes similar to those employed by the International Organization for Biological Control to rate pesticide toxicity to natural enemies of insect and mite pests. Selectivity of miticides was mostly based on toxicity to P ulmi, the major pest, versus toxicity to T pyri, the major predator, with some consideration of the two lesser species, A schlechtendali and Z mali. In most cases, our findings were in accord with other studies. Abamectin and clofentezine had favourable selectivity (more toxic to the two phytophagous mites than to T pyri). The higher recommended rate of pyridaben (450 g ha(-1)) and two rates of spirodiclofen (180 and 240 g ha(-1)) were neutral (equally toxic to pests and predators). The lower rate of pyridaben (216 g ha(-1)), dicofol, formetanate hydrochloride and propargite were unfavourably selective (more toxic to T pyri). A higher than recommended rate of pyridaben (2160 g ha(-1)) applied before bloom was disruptive--P ulmi-days after treatment were actually greater than with the untreated control. P ulmi resistance to dicofol and propargite were probable complicating factors in some of the orchard trials.
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In the 20th century an increasing number of pesticides, based on biocidal molecules, were the means for a substantial increase in food and fibre production and quality. Because of health and environmental concerns continued extensive use of such molecules is intensively debated and substitutes are often urgently required. Beside crop plant resistance, various biological control methods based on natural pest suppressing organisms are regarded as main alternatives. Several approaches and concepts also have been tested and commercial organism-based preparations are steadily increasing. However, further biotechnological efforts are required to give them status of being practical substitutes to pesticides. At present they are not comparable to pesticides in meeting efficacy, market and other expectations, but they still have a promising future, especially where genetically modified organisms can be used.
Entomopathogenic nematode species available in Europe were screened for their efficacy against both the root-feeding larvae and silk-feeding adults of the western corn rootworm, Diabrotica virgifera virgifera LeConte. Laboratory screening tests were aimed at the selection of candidate biological control agents for the management of this invasive alien pest in Europe. Steinernema glaseri, S. arenarium, S. abassi, S. bicornutum, S. feltiae, S. kraussei, S. carpocapsae and Heterorhabditis bacteriophora were studied to determine their virulence against third instar larvae and adults of D. v. virgifera in small-volume arenas (using nematode concentrations of 0.5, 0.8, 7.9 and 15.9 infective juveniles cm-2). All nematode species were able to invade and propagate in D. v. virgifera larvae, but adults were rarely infected. At concentrations of 7.9 and 15.9 cm-2, S. glaseri, S. arenarium, S. abassi and H. bacteriophora caused the highest larval mortality of up to 77%. Steinernema bicornutum, S. abassi, S. carpocapsae and H. bacteriophora appeared to have a high propagation level, producing 5970+/-779, 5595+/-811, 5341+/-1177 and 4039+/-1025 infective juveniles per larva, respectively. Steinernema glaseri, S. arenarium, S. feltiae, S. kraussei and H. bacteriophora were further screened at a concentration of 16.7 nematodes cm-2 against third instar larvae in medium-volume arenas (sand-filled trays with maize plants). Heterorhabditis bacteriophora, S. arenarium and S. feltiae caused the highest larval mortality with 77+/-16.6%, 67+/-3.5%, and 57+/-17.1%, respectively. In a next step, criteria for rating the entomopathogenic nematode species were applied based on results obtained for virulence and propagation, and for current production costs and availability in Europe. These criteria were then rated to determine the potential of the nematodes for further field testing. Results showed the highest potential in H. bacteriophora, followed by S. arenarium and S. feltiae, for further testing as candidate biological control agents.
In this paper, one investigates the dynamic behaviors of one-prey multi-predator model with Holling type II functional response by introducing impulsive biological control strategy (periodic releasing natural enemies at different fixed time). By using Floquet theorem and small amplitude perturbation method, it is proved that there exists an asymptotically stable pest-eradication periodic solution when the impulsive period is less than some critical value and permanence condition is established via the method of comparison involving multiple Liapunov functions. It is shown that multi-predator impulsive control strategy is more effective than the classical and single one.
Sequences, mixtures, rotations, and mosaics are potential strategies for using more than one pesticide to manage pest populations and for slowing the evolution of pesticide resistance. Results from theoretical models suggest that, under certain conditions, mixtures might be especially effective for resistance management. The assumptions of such models, however, are probably not widely applicable. Potential disadvantages associated with mixtures that are usually not considered in modeling studies include disruption of biological control, promotion of resistance in secondary pests, and intense selection for cross-resistance. Results from limited experimental work suggest that pesticide combinations do not consistently suppress resistance development. More thorough evaluation of tactics that seek to optimize benefits of more than one insecticide will require rigorous experiments with the particular pest and pesticide combinations. Because of the difficulty in generalizing results across systems and the potential negative effects of multiple insecticide use, emphasis on minimizing insecticide use is recommended.