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Degradation of the Cry1Ab protein within transgenic Bacillus thuringiensis corn tissue in the field.

Large quantities of Bacillus thuringiensis (Bt) corn plant residue are left in the field after harvest, which may have implications for the soil ecosystem. Potential impacts on soil organisms will also depend on the persistence of the Bt toxin in plant residues. Therefore, it is important to know how long the toxin persists in plant residues. In two field studies in the temperate corn-growing region of Switzerland we investigated degradation of the Cry1Ab toxin in transgenic Bt corn leaves during autumn, winter and spring using an enzyme-linked immunosorbent assay (ELISA). In the first field trial, representing a tillage system, no degradation of the Cry1Ab toxin was observed during the first month. During the second month, Cry1Ab toxin concentrations decreased to approximately 20% of their initial values. During winter, there was no further degradation. When temperatures again increased in spring, the toxin continued to degrade slowly, but could still be detected in June. In the second field trial, representing a no-tillage system, Cry1Ab toxin concentrations decreased without initial delay as for soil-incorporated Bt plants, to 38% of the initial concentration during the first 40 days. They then continued to decrease until the end of the trial after 200 days in June, when 0.3% of the initial amount of Cry1Ab toxin was detected. Our results suggest that extended pre- and post-commercial monitoring are necessary to assess the long-term impact of Bt toxin in transgenic plant residues on soil organisms.

Animals↗

Genetic transformation, recovery, and characterization of fertile soybean transgenic for a synthetic Bacillus thuringiensis cryIAc gene.

Somatic embryos of jack, a Glycine max (L.) Merrill cultivar, were transformed using microprojectile bombardment with a synthetic Bacillus thuringiensis insecticidal crystal protein gene (Bt cryIAc) driven by the 35S promoter and linked to the HPH gene. Approximately 10 g of tissue was bombarded, and three transgenic lines were selected on hygromycin-containing media and converted into plants. The recovered lines contained the HPH gene, but the Bt gene was lost in one line. The plasmid was rearranged in the second line, and the third line had two copies, one of which was rear-ranged. The CryIAc protein accumulated up to 46 ng mg-1 extractable protein. In detached-leaf bioassays, plants with an intact copy of the Bt gene, and to a lesser extent those with the rearranged copy, were protected from damage from corn earworm (Helicoverpa zea), soybean looper (Pseudoplusia includens), tobacco budworm (Heliothis virescens), and velvetbean caterpillar (Anticarsia gemmatalis). Corn earworm produced less than 3% defoliation on transgenic plants, compared with 20% on the lepidopteran-resistant breeding line GatIR81-296, and more than 40% on susceptible cultivars. Unlike previous reports of soybean transformation using this technique, all plants were fertile. To our knowledge, this is the first report of a soybean transgenic for a highly expressed insecticidal gene.

Animals↗

Genetics of pink bollworm resistance to Bacillus thuringiensis toxin Cry1Ac.

Laboratory selection increased resistance of pink bollworm (Pectinophora gossypiella) to the Bacillus thuringiensis toxin Cry1Ac. Three selections with Cry1Ac in artificial diet increased resistance from a low level to >100-fold relative to a susceptible strain. We used artificial diet bioassays to test F1 hybrid progeny from reciprocal crosses between resistant and susceptible strains. The similarity between F1 progeny from the two reciprocal crosses indicates autosomal inheritance of resistance. The dominance of resistance to Cry1Ac depended on the concentration. Resistance was codominant at a low concentration of Cry1Ac, partially recessive at an intermediate concentration, and completely recessive at a high concentration. Comparison of the artificial diet results with previously reported results from greenhouse bioassays shows that the high concentration of Cry1Ac in bolls of transgenic cotton is essential for achieving functionally recessive inheritance of resistance.

Animals↗

Synergistic efficacy of botanical blends with and without synthetic insecticides against Aedes aegypti and Culex annulirostris mosquitoes.

Increasing insecticide resistance requires strategies to prolong the use of highly effective vector control compounds. The use of combinations of insecticides with other insecticides and phytochemicals is one such strategy that is suitable for mosquito control. In bioassays with Aedes aegypti and Culex annulirostris mosquitoes, binary mixtures of phytochemicals with or without synthetic insecticides produced promising results when each was applied at a LC25 dose. All mixtures resulted in 100% mortality against Cx. annulirostris larvae within 24 h rather than the expected mortality of 50%. All mixtures acted synergistically against Ae. aegypti larvae within the first 24 h except for one mixture that showed an additive effect. We conclude that mixtures are more effective than insecticides or phytochemicals alone and that they enable a reduced dose to be applied for vector control potentially leading to improved resistance management and reduced costs.

Aedes↗

Evaluation of substituted oxime ethers for growth regulatory activity against Spodoptera litura (F.).

Insect growth regulatory activity (IGR) of fifty-two substituted oxime ethers were evaluated against an important polyphagous lepidopteran crop pest, Spodoptera litura (F.). A number of compounds produced symptoms comparable to exogenously applied juvenile hormone. Maximum IGR activity was exhibited by 4'-(2,6,6-trimethyl-2-cyclohexen- -yl)-3'-buten-2'(E)-ketoxime-N-O-alkyl ether with an ED50 (morphological) of 40 microg g(-1) body weight, compared to 20 microg g(-1) of JH III. Two more compounds namely 4'-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3'-buten-2'(Z)-ketoxime-N-O-methyl propyl ether (ED50 192 microg g(-1)) and 4'-(2,6,6-trimethyl-1-cyclohexen-1-yl)-3'-buten-2'(E)-ketoxime-N-O-pentyl ether (ED50 380 microg g(-1)) showed considerable IGR activity, whereas 4'-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3'-buten-2'(E)-ketoxime-N-O-pentyl ether was found to be toxic to the larvae (ED50 268 microg g(-1)). Three compounds used in this study were also synergised by piperonyl butoxide (PBO). The synergistic ratios were found in the range of 1.33 to 4.605. The ovicidal activity of the oxime ethers is not significant.

Animals↗

Suppression of Fusarium wilt by combining green compost and Trichoderma hamatum.

Fusarium wilts, caused by the fungus Fusarium oxysporum, are important diseases of horticultural and agricultural crops and lead to significant yield losses. The pathogen infects the roots and colonizes the vascular tissue, leading to wilting and finally death of the plant. The objective of this study was to investigate the efficacy of amendment of green compost and a Trichoderma hamatum strain against Fusarium wilt of radish. The substrate effects and the effect of a Trichoderma strain were tested in a potting soil bioassay. The tested composts lowered the disease level and had a positive influence on the plant yield (fresh weight and dry weight). Nothwithstanding, only a small dosis effect of the amendment was observed. In the presence of the tested Trichoderma hamatum strain no significant lower disease level was observed. Furthermore, a minor effect on plant yield (fresh weight and dry weight) was observed compared to the amendment with only green compost. Maybe the pathogenic Fusarium oxysporum strain and the Trichoderma strain competed for nutrients, iron could be a limiting factor. One possible approach to improve biological control may be the application of combinations of biocontrol agents.

Antibiosis↗

Assignment of the crystal toxin genes of the mosquitocidal bacterium, Bacillus thuringiensis israelensis to a specific plasmid.

Bacillus thuringiensis israelensis 4Q1-WT and the prepared mutants, 4Q1-72 and 4Q1-81 were bioassayed against Aedes caspius larvae. The strain -4Q1-WT, which contains all plasmid arrys and the strain 4Q1-72 which contains the 108 kb plasmid gave 100% mortality, while stain 4Q1-81 which contains no plasmids gave 0% mortality. Crystals from all tested strains were isolated, solubilized and characterized using PAGE to detect any homology or difference in crystal production, properties, and the relatedness to the plasmid profile.

Aedes↗

Methods for rearing Syngaster lepidus and Jarra phoracantha (Hymenoptera: Braconidae), larval parasitoids of the phloem-colonizing longhorned beetles Phoracantha semipunctata and P. recurva (Coleoptera: Cerambycidae).

Two species of cerambycid beetles that attack eucalypts, Phoracantha semipunctata (Fabricius) and P. recurva Newman, have been accidentally introduced from Australia into most regions of the world in which their hosts have been planted. The beetles cause extensive mortality in plantations and landscape plantings of the trees. Management programmes have focused on integration of silvicultural practices, host plant resistance and biological control. To rear and release natural enemies of the larval stages of the beetles in California, mass rearing protocols for continuous production of two species of parasitoids have been developed. The methods described represent the first long-term and large-scale techniques for mass rearing parasitoids of any wood-boring cerambycid species. In addition to providing large numbers of parasitoids for releases, the mass rearing effort has also provided large numbers of parasitoids for fundamental studies of their biology and behaviour.

Animals↗

Natural enemies of the maize cob borer, Mussidia nigrivenella (Lepidoptera: Pyralidae) in Benin, West Africa.

Mussidia nigrivenella Ragonot is a pest of maize cobs in West Africa. It significantly reduces maize yields and grain quality, with quantitative losses of 2-25%at harvest, and up to 10-15% indirect losses due to an increase in storage pest infestation levels. Infestation by M. nigrivenella also significantly increased the susceptibility of maize to Aspergillus flavus infection and subsequent aflatoxin contamination. Surveys conducted in different agro-ecological zones of Benin on cultivated and wild host plants during 1994-1997 revealed one egg parasitoid, three larval parasitoids and one pupal parasitoid attacking M. nigrivenella. Egg parasitism was scarce on all host plants sampled and in all four agro-ecological zones. Parasitism by larval and pupal parasitoids was usually less than 10%, and varied with host plant species. Both larval and pupal parasitoids were rare or absent in cultivated maize fields. The solitary chalcidid pupal parasitoid, Antrocephalus crassipes Masi, was the predominant species, contributing approximately 53% of the observed mortality. Logistic regression analysis indicated that this parasitoid was more prevalent on fruits of Gardenia spp. (Rubiaceae) than on the other host plant species including maize used by M. nigrivenella, and was most abundant between February and September. The differences in parasitoid diversity and parasitism between Benin and other regions suggest that there are opportunities for biological control through introduction of exotic parasitoids or using the 'new association' approach, which uses natural enemies of closely related host species that occupy similar ecological niches to the target pest.

Animals↗

Phytoecdysteroids: biological aspects.

Phytoecdysteroids are a family of about 200 plant steroids related in structure to the invertebrate steroid hormone 20-hydroxyecdysone. Typically, they are C27, C28 or C29 compounds possessing a 14alpha-hydroxy-7-en-6-one chromophore and A/B-cis ring fusion (5beta-H). In the present review, the distribution, biosynthesis, biological significance and potential applications of phytoecdysteroids are summarised.

Animals↗

Pesticide use, alternatives and workers' health in Cuba.

Cuba provides a unique example of a country that is actively implementing a program to reduce its dependence on pesticides. This paper addresses Cuba's current efforts to develop and implement alternatives to pesticides and legislation to limit exposure and protect workers in the interim. In 1980 Cuba embarked on a national program to utilize alternatives to chemical pest control. This three-part program includes expansion of knowledge of Cuban agro-ecology in order to implement cultural control practices; research and implementation on biological control of pests; and research on plant resistance and development of resistant crop varieties. To date, the program has enabled Cuba to reduce pesticide usage in sugar cane, citrus, tobacco, corn, and vegetable crops, among others. While alternatives to chemical pest control are being developed, the Cubans are paying special attention to regulating pesticide use and the safety of workers and members of the public exposed to toxic chemicals. In addition to the Resolution on Health and Safety (1967) and the Safety and Health Law (1978) which cover all workers, including Cuba's 250,000 agricultural workers, the Ministry of Public Health promulgated Resolution 335 in 1967. This resolution addresses requirements and administration of structural pest control, production, importation, transport and storage of pesticides, as well as requirements for worker contact with pesticides, pesticides for domestic use, aerial application of pesticides, and violations of the regulations. The paper concludes with a description of how the system works on the provincial level, as exemplified by Villa Clara, and the steps that have been taken to eliminate worker exposure to pesticides, to utilize pesticides which pose less of a hazard to workers, and to assure early detection of ill effects.

Agricultural Workers' Diseases↗