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Molecular cloning and expression of an alpha-amylase inhibitor from rye with potential for controlling insect pests.

Alpha-amylase inhibitors have important roles in plant defense mechanisms, particularly against insects, and several of these inhibitors have been expressed in different crops to increase their resistance to particular insects. In this work, we report the cloning and expression of a gene encoding for a new alpha-amylase inhibitor (BIII) from rye (Secale cereale) seeds. The BIII gene contains 354 nucleotides that encode for 118 amino acids sequence. A 313 bp fragment of the gene was expressed in Escherichia coli and resulted in a functional inhibitor that reduced the activity of alpha-amylases of larvae of the coleopteran pests Acanthoscelides obtectus, Zabrotess subfasciatus and Anthonomus grandis. In contrast, the inhibitor did not inhibit the activity of porcine pancreatic alpha-amylase. Although the amino acid sequence of BIII showed high identity with those of bifunctional inhibitors, the recombinant protein was unable to inhibit trypsin-like serine proteinases. The effects of recombinant BIII were evaluated in vivo against A. grandis. When first instar larvae were reared on an artificial diet containing four different concentrations of BIII, a reduction in larval weight and a mortality of 83% were observed at the highest concentration.

Amino Acid Sequence↗

Institutional pest management: current trends.

Insecticides and rodenticides will always be valuable tools in the pest control professional's battle against insect and vertebrate pests. But preventing and controlling pests in today's commercial and institutional facility has less to do with simply applying pesticides than it once did. Whether you are designing a pest control plan--or deciding which outside pest control contractor to hire--keep in mind that the best plan is not necessarily the one with the lowest price tag. Choose a service plan that includes pest exclusion, thorough inspections, and sanitation as first lines of defense against pests. Above all, make sure that the pest control plan you choose allows for plenty of communication between facility management and the people doing the actual pest control work.

Animals↗

Safety and advantages of Bacillus thuringiensis-protected plants to control insect pests.

Plants modified to express insecticidal proteins from Bacillus thuringiensis (referred to as Bt-protected plants) provide a safe and highly effective method of insect control. Bt-protected corn, cotton, and potato were introduced into the United States in 1995/1996 and grown on a total of approximately 10 million acres in 1997, 20 million acres in 1998, and 29 million acres globally in 1999. The extremely rapid adoption of these Bt-protected crops demonstrates the outstanding grower satisfaction of the performance and value of these products. These crops provide highly effective control of major insect pests such as the European corn borer, southwestern corn borer, tobacco budworm, cotton bollworm, pink bollworm, and Colorado potato beetle and reduce reliance on conventional chemical pesticides. They have provided notably higher yields in cotton and corn. The estimated total net savings to the grower using Bt-protected cotton in the United States was approximately $92 million in 1998. Other benefits of these crops include reduced levels of the fungal toxin fumonisin in corn and the opportunity for supplemental pest control by beneficial insects due to the reduced use of broad-spectrum insecticides. Insect resistance management plans are being implemented to ensure the prolonged effectiveness of these products. Extensive testing of Bt-protected crops has been conducted which establishes the safety of these products to humans, animals, and the environment. Acute, subchronic, and chronic toxicology studies conducted over the past 40 years establish the safety of the microbial Bt products, including their expressed insecticidal (Cry) proteins, which are fully approved for marketing. Mammalian toxicology and digestive fate studies, which have been conducted with the proteins produced in the currently approved Bt-protected plant products, have confirmed that these Cry proteins are nontoxic to humans and pose no significant concern for allergenicity. Food and feed derived from Bt-protected crops which have been fully approved by regulatory agencies have been shown to be substantially equivalent to the food and feed derived from conventional crops. Nontarget organisms exposed to high levels of Cry protein are virtually unaffected, except for certain insects that are closely related to the target pests. Because the Cry protein is contained within the plant (in microgram quantities), the potential for exposure to farm workers and nontarget organisms is extremely low. The Cry proteins produced in Bt-protected crops have been shown to rapidly degrade when crop residue is incorporated into the soil. Thus the environmental impact of these crops is negligible. The human and environmental safety of Bt-protected crops is further supported by the long history of safe use for Bt microbial pesticides around the world.

Animals↗

Endocrine strategies for the control of ectoparasites and insect pests.

The increasing knowledge about endocrine mechanisms in arthropods facilitates the biorational search for drugs against insect pests and parasites that interfere with arthropod hormone action. Juvenile hormone mimics have been successfully applied for about 20 years; however, resistance to juvenile hormone analogues has developed. The introduction of moulting hormone agonists, which compete for binding to the ecdysteroid receptor, is expected in the near future. Despite the considerable progress that has been achieved in peptide hormone research during the last few years, no successful insecticide is currently available, although comparisons of drugs for medical use demonstrate that in principle, successful interference with peptide hormone action is possible. The search for new drugs has been facilitated by advances in cell-culture techniques, which improve the development of suitable screening systems, and by progress in genetic engineering, which could be an important tool in the creation of new strategies for insect pest control.

Animals↗

Effects of plant protease inhibitors, oryzacystatin I and soybean Bowman-Birk inhibitor, on the aphid Macrosiphum euphorbiae (Homoptera, Aphididae) and its parasitoid Aphelinus abdominalis (Hymenoptera, Aphelinidae).

Transgenic plants expressing protease inhibitors (PIs) have emerged in recent years as an alternative strategy for pest control. Beneficial insects such as parasitoids may therefore be exposed to these entomotoxins either via the host or by direct exposure to the plant itself. With the objective of assessing the effects of PIs towards aphid parasitoids, bioassays using soybean Bowman-Birk inhibitor (SbBBI) or oryzacystatin I (OCI) on artificial diet were performed on Macrosiphum euphorbiae-Aphelinus abdominalis system. OCI significantly reduced nymphal survival of the potato aphid M. euphorbiae and prevented aphids from reproducing. This negative effect was much more pronounced than with other aphid species. On the contrary, SbBBI did not affect nymphal viability but significantly altered adult demographic parameters. Enzymatic inhibition assays showed that digestive proteolytic activity of larvae and adults of Aphelinus abdominalis predominantly relies on serine proteases and especially on chymotrypsin-like activity. Immunoassays suggested that OCI bound to aphid proteins and accumulated in aphid tissues, whereas SbBBI remained unbound in the gut. Bioassays using M. euphorbiae reared on artificial diets supplemented with both OCI and SbBBI showed a fitness impairment of Aphelinus abdominalis that developed on intoxicated aphids. However, only SbBBI was detected in parasitoid larvae, while no PI could be detected in adult parasitoids that emerged from PI-intoxicated aphids. The potential impact of PI-expressing plants on aphid parasitoids and their combined efficiency for aphid control are discussed.

Animals↗

Biorational agents--mechanism and importance in IPM and IRM programs for controlling agricultural pests.

Among the new approaches for controlling agricultural pests is the development of novel compounds affecting specific processes in insects such as chitin synthesis inhibitors, juvenile hormone mimics and ecdysone agonists. In addition, efforts have been made to develop compounds acting selectively on groups of insects by inhibiting or enhancing biochemical sites such as respiration (diafenthiuron), the nicotinyl acetylcholine receptors (imidacloprid and acetamiprid), the GABA receptors (avermectins), the salivary glands of sucking pests (pymetrozine) and others. Among the most recent novel insecticides with selective properties are novaluron, thiamethoxam, emamectin and spinosad. Novaluron (Rimon) is a novel chitin synthesis inhibitor that acts by both ingestion and contact. It is a powerful suppressor of lepidopteran larvae such as Spodoptera littoralis and Helicoverpa armigera (by ingestion) and of whiteflies such as Bemisia tabaci and Trialeurodes vaporariorum (by contact). Thiamethoxam (Actarn), a novel neonicotinoid acts specifically on aphids and whiteflies. Emamectin (Proclaim), an avermectin derivative acts on GABA receptor affecting diversity of insects such as mites, lepidopterans and thrips. Spinosad (Tracer) seems to act on both acetylcholine and GABA receptors affecting diversity of insect species and is considered an important agent for controlling the western flower thrips.

Animals↗

Production efficiency of loblolly pine and sweetgum in response to four years of intensive management.

We tested the hypothesis that productivity of intensively managed loblolly pine (Pinus taeda L.) and sweetgum (Liquidambar styraciflua L.) stands is dependent not only on leaf area, but also on foliar photosynthetic rate. Effects of irrigation (irrigation treatment), irrigation with a fertilizer solution (fertigation treatment), and fertigation + pest control (loblolly pine only; fertigation + pest control treatment) on leaf physiology and growth were compared with control plots during the third and fourth growing seasons. Complete weed control was maintained on all plots. Aboveground net primary productivity of loblolly pine and sweetgum increased from 16.3 to 40.5 Mg ha(-1) and from 4.2 to 23.9 Mg ha(-1), respectively, in response to the most intensive treatment. Relative to the control treatment, neither fertigation of sweetgum nor fertigation + pest control of loblolly pine had a significant or consistent influence on foliar N concentration, quantum yield, carboxylation efficiency, net photosynthesis, stomatal conductance, or production efficiency (increment in woody biomass per unit leaf area). Irrigation increased predawn leaf water potential and photosynthesis of loblolly pine, but it had no effect on production efficiency. Leaf area was the predominant determinant of maximum productivity in these rapidly growing plantations.

Forestry↗

Residential pesticide use during pregnancy among a cohort of urban minority women.

Residential pesticide use is widespread in the United States. However, data are limited specific to use among minority populations. Nor are data available on the extent of pesticide exposure resulting from residential use during pregnancy. We have gathered questionnaire data on pesticide use in the home during pregnancy from 316 African-American and Dominican women residing in northern Manhattan and the South Bronx. Additionally, 72 women underwent personal air monitoring for 48 hr during their third trimester of pregnancy to determine exposure levels to 21 pesticides (19 insecticides and 2 fungicides). Of the women questioned, 266 of 314 (85%) reported that pest control measures were used in the home during pregnancy; 111 of 314 (35%) reported that their homes were sprayed by an exterminator, and of those, 45% said the spraying was done more than once per month. Most (>or= 90%) of the pesticide was used for cockroach control. Use of pest control measures increased significantly with the level of housing disrepair reported. Of the women monitored, all (100%) had detectable levels of three insecticides: the organophosphates diazinon (range, 2.0-6,010 ng/m3) and chlorpyrifos (range, 0.7-193 ng/m3) and the carbamate propoxur (range, 3.8-1,380 ng/m3), as well as the fungicide o-phenylphenol (range, 5.7-743 ng/m3). We also frequently detected the following four insecticides (47-83% of samples) but at lower concentrations: the pyrethroid trans-permethrin, piperonyl butoxide (an indicator of exposure to pyrethrins), and the organochlorines 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane and chlordane. Thirty percent of the women had detectable levels of all eight pesticides. Exposures were generally higher among African Americans than among Dominicans. We detected other pesticides in <or= 10% of samples. Results show widespread prenatal pesticide use among minority women in this cohort. Diazinon exposures for some women may have exceeded health-based levels, and our findings support recent federal action to phase out residential use of this insecticide.

Adult↗

Prevention and control of pests and diseases.

A well-controlled growth environment with plants that are not unduly stressed is essential for Arabidopsis molecular biology research. Even if they do not kill the plants outright, insect pests and microbial pathogens can cause subtle changes in gene expression or plant metabolism that affect experimental results. Therefore, regular scouting for infestations, frequent cleaning of plant growth areas, proper disposal of dead or diseased plant material, and controlled access to the greenhouses or growth chambers will help to make experiments more reproducible. Powdery mildew, a fungal pathogen, and arthropod pests, including aphids, thrips, fungus gnats, and spider mites, are the most common greenhouse problems. Biological control methods such as parasitoid wasps and Bacillus thuringiensis crystal toxin can be used to contain some insect infestations. However, if an infestation gets out of hand despite reasonable precautions, insecticide or fungicide spraying by a licensed applicator may be necessary. Bacterial and viral infections of Arabidopsis, though they do occur, tend to be less common and can usually be controlled by maintaining optimal growth conditions and promptly disposing of dead or diseased plant material.

Animals↗

Ecdysone agonists: mechanism and importance in controlling insect pests of agriculture and forestry.

Molting is the result of the expression of a cascade of genes that is sequentially both up and down-regulated by the molting hormone, 20-hydroxyecdysone (20E), which is secreted as a pulse during each instar. Benzoyl hydrazine analogs of 20E act like the native molting hormone at the molecular level by binding with the ecdysone receptor complex and transactivating a succession of molt initiating transcription factors that, in turn, induce the expression of a group of molt-related genes. As a result of the expression of these up-regulated genes, the larva undergoes apolysis and head capsule slippage and takes on the appearance of the pharate larva. However, unlike 20E, which is cleared at this juncture, allowing the down-regulated genes to be expressed, these synthetic analogs bind strongly to the receptors and remain in place and repress all the down-regulatory genes such as the ones necessary for cuticle elaboration, sclerotization, and ecdysis resulting in a developmental arrest in this state. As a result, the treated larva goes into a precocious incomplete molt that is lethal. Two of the analogs, tebufenozide and methoxyfenozide, are lepidopteran specific and have good control potential for open feeding larvae that ingest this material while a third one, halofenozide, acts on coleopteran larvae. Since they specifically act through an insect receptor complex, they have little or no effect on non-target species, making them environmentally attractive pest control agents. Some insects, however, show resistance to these analogs and this could be, inter alia, due to an ATP Binding Cassette Transporter like system that selectively pumps out the analogs.

ATP-Binding Cassette Transporters↗

Control of pest blackflies (Diptera:Simuliidae) along the Orange River, South Africa: 1990-1995.

The efficacy of Bacillus thuringiensis var. israelensis (B.t.i.) and temephos in controlling the pest blackfly Simulium chutteri Lewis along the middle Orange River between 1990 and 1995, was assessed. Larvicides were applied by helicopter to rapids and riffles between Hopetown and Onseepkans, a river distance of 807 km. Larvicidal efficacy was based on the change in larval abundance at selected sites before and after each treatment. The success of the control programme was assessed independently by local farmers, who ranked adult blackfly annoyance on a 4-point scale. Before treatment, blackfly annoyance showed consistent peaks in spring, and sometimes in autumn, and levels were unacceptably high for between 17 and 36 weeks of the year. After treatment started, blackfly annoyance levels were reduced significantly. The number of annual treatments necessary to reduce blackfly annoyance to acceptable levels was highly variable (3-13), and depended on river conditions, as well as the efficacy and timing of each treatment. During low-flow conditions (< 50 m3/s), applications became increasingly difficult in braided sections of the river, and dosage calculations were inaccurate because of local abstraction and return flows. Both larvicides worked well in winter (water temperature 11-13 degrees C). Control of the spring outbreak can be planned well in advance, with the first treatment starting in mid July. A flexible protocol is required to control outbreaks at other times of the year. We recommended the use of B.t.i. for most applications, with increased dosages during algal blooms (> 1500 cells/ml). The use of temphos in the Orange River should be considered only during algal blooms or when flows exceed 300 m3/s. We conclude that helicopter application of larvicides is an effective method or controlling blackflies, along the middle Orange River.

Animals↗