The effect of sucrose content of diets on susceptibility to granulosis virus disease in Pieris brassicae.
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To conduct laboratory experiments aimed at quantifying secondary acquisition of fungal conidia by western flower thrips (Frankliniella occidentalis), an efficient assay technique using Beauveria bassiana as the model fungus was developed. Various application protocols were tested and it was determined that the percent mortality did not vary among protocols. Peak mortality of second-instar nymphs, under constant exposure to conidia, occurred 5 days post-inoculation. Second-instar thrips that were exposed to conidia within 24 h of the molt to second instar were more susceptible to Beauveria bassiana than thrips exposed after times greater than 24 h post-molt. Conidia efficacy, which was monitored at 24 h intervals, did not differ significantly within 72 h. A test of the final bioassay system was conducted in a series of assays aimed at determining the LD50 of B. bassiana technical powder against second-instar western flower thrips. It was determined that B. bassiana (strain GHA) is highly effective at very low doses (LD50 of 33-66 conidia/insect).
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We describe the isolation, cloning and expression of a 2.8 kb promoter fragment of the Blec4 gene from pea (Pisum sativum cv. Alaska) and demonstrate that it is capable of directing the expression of the beta-glucuronidase coding region to the developing epidermal tissue of vegetative and floral shoot apices of transgenic alfalfa (Medicago sativa cv. RegenSY). The Blec4 promoter represents a useful tool with which to target the expression of foreign genes to the epidermal layer of actively growing shoots. The activity of the Blec4 promoter in the epidermis of the shoot apex makes it particularly suitable for genetically engineering defense against insects and diseases that attack the growing shoot apex.
The impact of relative humidity (RH) on the infective potential of the isolate Bb INRA 297 of Beauveria bassiana (Bals.) Vuillemin (Deuteromycotina: Hyphomycetes) against first instar nymphs of Rhodnius prolixus Stål. (Hemiptera: Reduviidae) was determined. Fungus-treated insects were exposed to RHs ranged from 75 to 100% at 25 degrees C. Results clearly showed a threshold of humidity at ca. 96% for high and rapid mortality. After initial exposure to increasing periods of 97% (4, 8, 16, 24, 36 and 48 h) and subsequent transfer to constant lower RHs (43, 53, 75 and 86%) at a constant 25 degrees C, an incubation of at least 48 h at 97% RH was necessary to kill all insects. On changing RHs of 97/75% and different regimes of temperature (15/28 degrees C, 20/25 degrees C, 25/28 degrees C, and 25/35 degrees C), at least 72 h of initial exposure at 97% RH for the 15/28 degrees C regime, 48 h for the 20 degrees/25 degrees C and 25/28 degrees C regimes and 36 h for 25/35 degrees C were needed to kill all insects over a 6-day incubation time. Delayed exposure to favorable moisture condition (97% RH), significantly affected infection for up to a 3-day delay within the various temperature-humidity regimes tested.
Pauesia juniperorum (Starý), a solitary endoparasitoid which attacks several Cinara spp. in the subgenus Cupressobium, was introduced into Africa for the control of the cypress aphid Cinara cupressivora Watson & Voegtlin (previously identified as Cinara cupressi (Buckton)) and is now established. Data on its biology including a comparison of populations from different hosts and geographical locations are presented. From the UK this consisted of populations reared from Cinara cupressivora, Cinara fresai Blanchard and Cinara juniperi De Geer, and from France, Cinara ?fresai. Between different populations, the duration of development varied within a narrow range (15.5-18.1 days) at 22 degrees C. Data on duration of development over a range of temperatures (15-26 degrees C) for two populations, from the UK and France ranged between 14.0-31.4 days for males and 14.7-32.8 days for females. The threshold temperature for development was 6.2 degrees C and 4.1 degrees C for the populations from the UK and France, respectively. The mean egg load for different populations ranged between 46.4 and 78.5 eggs. The host species had a significant effect on egg load which was directly related to size of the respective host aphids. The lowest egg load was recorded on specimens reared from C. cupressivora which was the smallest aphid. The population from the UK had a higher intrinsic rate of increase (0.232) than the one from France (0.207).
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The potential of using phytochemicals from leaves of wild tomato for controlling the two-spotted spider mite, Tetranychus urticae Koch, is explored in this study as a promising alternative to the use of synthetic pesticides. Wild tomato accessions of Lycopersicon hirsutum plants that are not consumed by humans were planted under greenhouse conditions for mass production of leaves. Crude extracts from leaves of three accessions of L. hirsutum, six accessions of L. hirsutum f. glabratum, and one accession each of L. pennellii and L. pimpinellifolium were prepared in chloroform, ethanol and hexane. Two spider mite bioassays, one a measure of antibiosis and the other a measure of repellency, were utilized to determine the acaricidal performance of the crude extracts. The bioassay for antibiosis was a 6-h no-choice test. The bioassay for repellency utilized a ring bioassay. Chloroform leaf extracts of L. hirsutum f. glabratum accessions (PI-251304, PI-134417, PI-134418, and PI-126449) exhibited greatest antibiotic activity on two-spotted spider mites; the hexane extracts exhibited greatest repellency. Extracts from PI-251304, PI-126449, PI-134417, and PI-134418 were especially lethal (chloroform) or repellent (hexane). We investigated differences in chemical composition of the crude leaf extracts that may explain the observed differences in mortality and repellency among the different accessions. Major chemical compounds (alpha -curcumene, alpha -zingiberene, trans-caryophyllene, 2-undecanone, and 2-tridecanone) known to have pesticidal efficacy were detected and quantified in the crude leaf extracts using a gas chromatograph (GC) equipped with a mass spectrometer (GC/MS). Lethality of extracts was mainly associated with the presence of high concentrations of 2-tridecanone; repellency of extracts was mainly associated with the presence of trans-caryophyllene. Leaf extracts of L. hirsutum f. glabratum accessions that contain significant quantities of 2-tridecanone and/or trans-caryophyllene could be useful for managing populations of spider mites, which could reduce reliance on synthetic pesticides.
Beauveria bassiana (Balsamo) Vuillemin (isolate CG306) was tested on 3rd instars of 9 Triatoma spp., 4 Rhodnius spp., 2 Panstrongylus spp. and Dipetalogaster maxima (Uhler) at 25 degrees C and 50% RH. Quantitative sporulation of the fungus on cadavers was studied at 25 degrees C and 97% RH. Mortality, estimates of survival time, and conidial production on cadavers differed significantly among the genera and species tested. Panstrongylus herreri Wygodzinsky, Dipetalogaster maxima, Triatoma picturata Usinger, Rhodnius robustus Larrousse, Rhodnius prolixus Stål, Triatoma infestans (Klug), and Triatoma brasiliensis Neiva were most susceptible to fungal infection. Most conidia per cadaver were produced on Triatoma williami Galvão, Souza & Lima and Triatoma lecticularia (Stål). Results indicate that B. bassiana (CG306) might be a candidate for control of important or potential vectors of Trypanosoma cruzi using the fungus as a bioinsecticide.
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Fumonisins (FB) are mycotoxins found in (italic)Fusarium verticillioides-infected maize grain worldwide. Attention has focused on FBs because of their widespread occurrence, acute toxicity to certain livestock, and their potential carcinogenicity. FBs are present at low levels in most field-grown maize but may spike to high levels depending on both the environment and genetics of the host plant. Among the strategies for reducing risk of FB contamination in maize supplied to the market, development and deployment of Fusarium ear mold-resistant maize germplasm is a high priority. Breeding for increased ear mold tolerance and reduced mycotoxin levels is being practiced today in both commercial and public programs, but the amount of resistance achievable may be limited due to complicated genetics and/or linkage to undesirable agronomic traits. Molecular markers can be employed to speed up the incorporation of chromosomal regions that have a quantitative effect on resistance (quantitative trait loci). Transgenic approaches to ear mold/mycotoxin resistance are now feasible as well. These potentially include genetically enhanced resistance to insect feeding, increased fungal resistance, and detoxification/prevention of mycotoxins in the grain. An example of the first of these approaches is already on the market, namely transgenic maize expressing Bacillus thuringiensis (Bt) toxin, targeted to the European corn borer. Some Bt maize hybrids have the potential to reduce FB levels in field-harvested grain, presumably through reduced feeding of Bt-susceptible insects in ear tissues. However, improved ear mold resistance per se is still an important goal, as the plant will still be vulnerable to noninsect routes of entry to (italic)Fusarium. A second approach, transgene-mediated control of the ability of Fusarium to infect and colonize the ear, could potentially be achieved through overexpression of specific antifungal proteins and metabolites, or enhancement of the plant's own defense systems in kernel tissues. This has not yet been accomplished in maize, although promising results have been obtained recently in other monocots versus other fungal and bacterial pathogens. Achieving reproducible and stable enhanced ear mold resistance under field conditions will be immensely challenging for biotechnologists. A third approach, transgene strategies aimed at preventing mycotoxin biosynthesis, or detoxifying mycotoxins in planta, could provide further protection for the grower in environments where FBs present a risk to the crop even when the maize is relatively resistant to Fusarium mold. In one example of such a strategy, enzymes that degrade FBs have been identified in a filamentous saprophytic fungus isolated from maize, and corresponding genes have been cloned and are currently being tested in transgenic maize.
Dispersal of neonate European corn borers, Ostrinia nubilalis (Hübner), in seed mixtures of transgenic corn expressing Cry1Ab protein (Bt+) and nontransgenic corn (Bt-) was evaluated in a 2-yr field study. The main objective was to determine if larval dispersal limits the effectiveness of seed mixtures as a resistance management strategy. Mixtures evaluated included (1) all Bt+ plants, (2) every fifth plant Bt- with remaining plants Bt+, (3) every fifth plant Bt+ with remaining plants Bt-, and (4) all Bt- plants. The transformation events MON 802 (B73 BC1F2 x Mol7) and MON 810 (B73 BC1F1 x Mo17), which express the Cry1Ab endotoxin isolated from Bacillus thuringiensis subsp. kurstaki, were used as the sources of Bt+ seed in 1994 and 1995, respectively (YieldGard, Monsanto, St. Louis, MO). At corn growth stage V6-V8, subplots within each mixture (15-20 plants each) were infested so that every fifth plant in mixtures 1 and 4, every Bt- plant in mixture 2, and every Bt+ plant in mixture 3 received two egg masses. Larval sampling over a 21-d period indicated increased neonate dispersal off of Bt+ plants, reduced survival of larvae that dispersed from Bt+ plants to Bt- plants, and a low incidence of late-instar movement from Bt- plants to Bt+ plants. Computer simulations based on mortality and dispersal estimates from this study indicate that seed mixtures will delay the evolution of resistant European corn borer populations compared with uniform planting of transgenic corn. However, resistant European corn borer populations likely will develop faster in seed mixes compared with separate plantings of Bt and non-Bt corn.
Leaves of the tropical legume Tephrosia candida DC deterred feeding by adults of the Diaprepes root weevil, Diaprepes abbreviatus (L.), compared with leaves of Citrus macrophylla Wester, a common citrus rootstock, or T. vogelii Hook. f. When larvae were placed in pots containing plants of the three species for 28 d in a growth chamber, larval survival and weight gain were significantly reduced in pots containing plants of T. candida compared with larvae in pots with C. macrophylla or T. vogelii. Diet incorporation of lyophilized roots of T. candida into an artificial diet increasingly inhibited the growth of larvae and increased larval mortality with increased concentration of roots, whereas roots from C. macrophylla or T. vogelii had no effect compared with the diet-only control. T. candida, but not T. vogelii, contains at least one constituent that acts as an antifeedant toward adult D. abbreviatus and as a toxicant toward larvae. No antifeedant effect of roots of T. candida toward larvae was observed in no-choice pot tests or in a diet incorporation bioassay. In pots, larval feeding damage to roots of T. candida was evident. In the diet incorporation assay, 97% of larvae survived 29 d on a diet of cellulose powder (a nutritionally inert filler) despite losing weight. We conclude that decreased survival and weight gain of larvae-fed fresh or lyophilized roots of T. candida were the result of ingestion of a toxicant and not deterrence from feeding.
Bacterial symbionts may be used as vehicles for expressing foreign genes in arthropods. Expression of selected genes can render an arthropod incapable of transmitting a second microorganism that is pathogenic for humans and is an alternative approach to the control of arthropod-borne diseases. We discuss the rationale for this alternative approach, its potential applications and limitations, and the regulatory concerns that may arise from its use in interrupting disease transmission in humans and animals.
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