Biomedical subjects

D Bourguet

Publications and source records attributed to D Bourguet.

At least 19 recordsLinked to original sources

PubMed2000

Host-plant diversity of the European corn borer Ostrinia nubilalis: what value for sustainable transgenic insecticidal Bt maize?

The strategies proposed for delaying the development of resistance to the Bacillus thuringiensis toxins produced by transgenic maize require high levels of gene flow between individuals feeding on transgenic and refuge plants. The European corn borer Ostrinia nubilalis (Hübner) may be found on several host plants, which may act as natural refuges. The genetic variability of samples collected on sagebrush (Artemisia sp.), hop (Humulus lupulus L.) and maize (Zea mays L.) was studied by comparing the allozyme frequencies for six polymorphic loci. We found a high level of gene flow within and between samples collected on the same host plant. The level of gene flow between the sagebrush and hop insect samples appeared to be sufficiently high for these populations to be considered a single genetic panmictic unit. Conversely, the samples collected on maize were genetically different from those collected on sagebrush and hop. Three of the six loci considered displayed greater between-host-plant than within-host-plant differentiation in comparisons of the group of samples collected on sagebrush or hop with the group of samples collected on maize. This indicates that either there is genetic isolation of the insects feeding on maize or that there is host-plant divergent selection at these three loci or at linked loci. These results have important implications for the potential sustainability of transgenic insecticidal maize.

Adaptation, Physiological↗

PubMed2000

Gene flow in the European corn borer Ostrinia nubilalis: implications for the sustainability of transgenic insecticidal maize.

Strategies proposed for delaying resistance to Bacillus thuringiensis toxins expressed by transgenic maize require intense gene flow between individuals that grew on transgenic and on normal (referred to as refuges) plants. To investigate gene flow in the European corn borer, Ostrinia nubilalis (Hübner), the genetic variability at 29 sampled sites from France was studied by comparing allozyme frequencies at six polymorphic loci. Almost no deviations from Hardy-Weinberg expectations occurred, and a high stability of allelic distribution was found among samples collected in the same site over two or three different generations, indicating a high stability of the genetic structure over time. The overall genetic differentiation was low at the region and whole country level, suggesting a high and homogeneous gene flow. These results are discussed in relation to the sustainability of transgenic insecticidal maize.

Animals↗

PubMed2000

Factors affecting the genetic load in Drosophila: synergistic epistasis and correlations among fitness components.

Two factors that can affect genetic load, synergistic epistasis and sexual selection, were investigated in Drosophila melanogaster. A set of five chromosomal regions containing visible recessive mutations were put together in all combinations to create a full set of 32 homozygous lines fixed for different numbers of known mutations. Two measures of fitness were made for each line: productivity (a combined measure of fecundity and egg-to-adult survivorship) and competitive male mating success. Productivity, but not male mating success, showed a pattern of strong average synergistic epistasis, such that the log fitness declined nonlinearly with increasing numbers of mutations. Synergistic epistasis is known to reduce the mutation load. Both fitness components show some positive and some negative interactions between specific sets of mutations. Furthermore, alleles with deleterious effects on productivity tend to also diminish male mating success. Given that male mating success can affect relative fitness without changing the mean productivity of a population, these additional effects would lead to lower frequencies and lower fixation rates of deleterious alleles without higher costs to the mean fitness of the population.

Animals↗

PubMed2000

Population genetics of Chrysomela tremulae: a first step towards management of transgenic Bacillus thuringiensis poplars Populus tremula x .P. tremuloides.

Many strategies have been proposed for delaying the development of insect resistance to Bacillus thuringiensis (Bt). The current paradigm for Bt resistance management is the high dose-refuge strategy. For this strategy to be successful: (i) heterozygotes must be killed in treated areas, (ii) resistant alleles must be rare (frequency < 10-3), and (iii) there must be a high level of gene flow between populations to ensure random mating. We studied gene flow within and between populations with a view to managing the resistance of Chrysomela tremulae (Coleoptera: Chrysomelidae) to new transgenic, highly toxic poplars expressing a synthetic Bt gene. In this study, we assessed the extent of gene flow in C. tremulae within and between 16 sites in France and Belgium, using allozyme markers. We found a high level of genetic variability in C. tremulae, with a mean of 0.206 +/- 0.16. There were no obvious limitations to gene flow between populations of C. tremulae over large geographical distances (several hundreds of kilometres). Nevertheless, a very low level of genetic differentiation was observed between a site located in the south of France and the sampled sites from the Centre region.

Animals↗

PubMed2000

Insecticide resistance and dominance levels.

Dominance has been assessed in different ways in insecticide resistance studies, based on three phenotypic traits: the insecticide concentration required to give a particular mortality (DLC), mortality at a particular insecticide dose (DML), and fitness in treated areas (DWT). We propose a general formula for estimating dominance on a scale of 0 to 1 (0 = complete recessivity and 1 = complete dominance). DLC, DML, and DWT are not directly related and their values depend on genetic background and environmental conditions. We also show that pest management strategies can have the consequence to increase DWT via the selection of dominance modifiers. Studies on resistance to Bacillus thuringiensis toxins provide the ultimate example of the complexity of the definition of the concept of dominance. Almost all studies have focused on calculation of DLC, which provides little information about the efficiency of pest management programs. For instance, one assumption of the high dose/refuge strategy is that Bacillus thuringiensis resistance must be effectively recessive (i.e., DML must be close to zero). However, DWT, rather than DML, is relevant to the resistance management strategy. Therefore, we strongly suggest that the time has come to focus on fitness dominance levels in the presence and absence of insecticide.

Animals↗

PubMed1999

Tracking the evolution of insecticide resistance in the mosquito Culex pipiens.

The evolution of pesticide resistance provides some of the most striking examples of darwinian evolution occurring over a human life span. Identification of resistance alleles opens an outstanding framework in which to study the evolution of adaptive mutations from the beginning of pesticide application, the evolution of interactions between alleles (dominance) or between loci (epistasis). Here we show that resistance alleles can also be used as markers to dissect population processes at a microevolutionary scale. We have focused on the antagonistic roles of selection and migration involved in the dynamics of local adaptation with reference to allelic frequencies at two resistance loci in the mosquito Culex pipiens. We find that their frequencies follow an annual cycle of large amplitude (25%), and we precisely unravel the seasonal variation of migration and selection underlying this cycle. Our results provide a firm basis on which to devise an insecticide treatment strategy that will better control the evolution of resistance genes and the growth of mosquito populations.

Alleles↗

PubMed1999

The evolution of dominance.

The evolution of dominance has been subject to intensive debate since Fisher first argued that modifiers would be selected for if they made wild-type alleles more dominant over mutant alleles. An alternative explanation, put forward by Wright, is that the commonly observed dominance of wild-type alleles is simply a physiological consequence of metabolic pathways. Wright's explanation has gained support over the years, largely ending the debate over the general recessivity of deleterious mutations. Nevertheless there is reason to believe that dominance relationships have been moulded by natural selection to some extent. First, the metabolic pathways are themselves products of evolutionary processes that may have led them to be more stable to perturbations, including mutations. Secondly, theoretical models and empirical experiments suggest that substantial selection for dominance modifiers exists during the spread of adaptive alleles or when a polymorphism is maintained either by overdominant selection or by migration-selection balance.

Animals↗

PubMed1999

A new mechanism conferring unprecedented high resistance to chlorpyrifos in Culex pipiens (Diptera: Culicidae).

The cause of high resistance to chlorpyrifos observed in Tunisian Culex pipiens (L.) was investigated by comparing a Tunisian strain G (> 10,000-fold resistance), a French strain T (approximately 50-fold resistance), and a susceptible reference strain S. Strains G and T had the same level of propoxur resistance (approximately 1,000-fold) and were homozygous for an autosomal propoux-insensitive acetylcholinesterase (AChE-1). In G and T strains, as well as in the offspring of different F1s and backcrosses using these F1s and the S strain, the effect of DEF and Pb synergists on chlorpyrifos resistance was low or absent, indicating that increased detoxification by enzymes inhibited by these chemicals had a minor role. Chlorpyrifos resistance in the G strain was caused by a major gene (or group of genes) tightly linked to the Ace-1 gene (coding AChE-1 enzyme). The possibility of allelism between this gene and the Ace-1R allele present in the T strain was rejected by showing that AChE-1 inhibition by chlorpyrifos-oxon was not different between G and T mosquitoes.

Acetylcholinesterase↗

PubMed1998

A sex-linked Ace gene, not linked to insensitive acetylcholinesterase-mediated insecticide resistance in Culex pipiens.

An acetylcholinesterase (AChE) gene, Ace.x, showing 93% identity of deduced amino acid sequence to Anopheles stephensi Ace has been cloned from a Culex pipiens strain homozygous for insensitive AChE (iAChE) mediated insecticide resistance. DNA sequence of genomic DNA clones identified exons 2-5. RFLP of six clones indicated four possible alleles. Linkage analysis located Ace.x to chromosome I, less than 0.8 centimorgans from the sex locus, whereas the locus conferring resistance was 2.0 centimorgans from plum-eye on chromosome II. Ace.1 coding for AChE1, which is associated with resistance, is therefore autosomal. We propose that Ace.x is the recently postulated Ace.2 coding for the biochemically distinct AChE2, which is not associated with resistance.

Acetylcholinesterase↗

PubMed1998

Evaluating gene flow using selected markers: a case study.

The extent to which an organism is locally adapted in an environmental pocket depends on the selection intensities inside and outside the pocket, on migration, and on the size of the pocket. When two or more loci are involved in this local adaptation, measuring their frequency gradients and their linkage disequilbria allows one to disentangle the forces-migration and selection-acting on the system. We apply this method to the case of a local adaptation to organophosphate insecticides in the mosquito Culex pipiens pipiens in southern France. The study of two different resistance loci allowed us to estimate with support limits gene flow as well as selection pressure on insecticide resistance and the fitness costs associated with each locus. These estimates permit us to pinpoint the conditions for the maintenance of this pocket of adaptation as well as the effect of the interaction between the two resistance loci.

Animals↗

PubMed1998

The acetylcholinesterase gene Ace: a diagnostic marker for the Pipiens and Quinquefasciatus forms of the Culex pipiens complex.

The taxonomy of the Culex pipiens complex remains a controversial issue in mosquito systematics. Based on morphologic characters, 2 allopatric taxa are recognized, namely Cx. pipiens (including the form "molestus") in temperate areas and Cx. quinquefasciatus in tropical areas. Here we report on variability at the nucleotide level of an acetylcholinesterase gene in several strains and natural populations of this species complex. Few polymorphisms were found in coding regions within a subspecies but many polymorphisms were observed between subspecies in noncoding regions. We describe a method based on a restriction enzyme polymorphism in polymerase chain reaction-amplified DNA, in which the presence or absence of one restriction site discriminates Cx. pipiens, Cx. quinquefasciatus, and their hybrids. This technique reliably discriminates mosquitoes from more than 30 worldwide strains or populations. Polymerase chain reaction amplification of specific alleles may also be a useful tool for characterizing specific alleles of each sibling taxon.

Acetylcholinesterase↗

PubMed1997

Analysis of molecular forms and pharmacological properties of acetylcholinesterase in several mosquito species.

Two acetylcholinesterases (AChE1 and AChE2) have recently been characterized in the common mosquito Culex pipiens. This situation appeared to be an exception among insects, where only one acetylcholinesterase gene had previously been repeatedly reported. In the present study, acetylcholinesterase was studied in five mosquito species: Aedes aegypti, Anopheles gambiae, Anopheles stephensi, Culiseta longeareolata and Culex hortensis, in order to test whether or not two different acetylcholinesterase enzymes could be detected as occurs in C. pipiens. Molecular forms and catalytic properties of the enzyme show that only one enzyme species was detected in the five species. This suggests that a duplication of a single locus Ace probably occurred recently in the phylogeny tree leading to C. pipiens, and produced two distinct acetylcholinesterases: AchE1 and AChE2.

Acetylcholinesterase↗

PubMed1997

Pleiotropy of adaptive changes in populations: comparisons among insecticide resistance genes in Culex pipiens.

Resistance to toxicants is a convenient model for investigating whether adaptive changes are associated with pleiotropic fitness costs. Despite the voluminous literature devoted to this subject, intraspecific comparisons among toxicant resistance genes are rare. We report here results on the pleiotropic effect on adult survival of Culex pipiens mutants involved in the same adaptation: the resistance to organophosphorus insecticides. This field study was performed in southern France where four resistance genes sequentially appeared and increased in frequency in response to intense insecticide control. By repeated sampling of overwintering females through winter, we analysed the impact of each of three resistance genes on adult survival. We showed that (i) the most recent gene seems to be of no disadvantage during winter, (ii) the oldest affects survival in some environmental conditions, and (iii) the third induces a constant, severe and dominant survival cost. Such variability is discussed in relation to the physiological changes involved in resistance.

Acetylcholinesterase↗

PubMed1997

Variation of dominance of newly arisen adaptive genes.

Newly arisen adaptive alleles such as insecticide resistance genes represent a good opportunity to investigate the theories put forth to explain the molecular basis of dominance and its possible evolution. Dominance levels of insecticide resistance conferred by insensitive alleles of the acetylcholinesterase gene were analyzed in five resistant strains of the mosquito Culex pipiens. Dominance levels were found to differ between strains, varying from partial recessivity to complete dominance. This variation was not explained by differences in catalytic properties of the enzyme, since four of the five resistant strains had identical inhibition properties for the insensitive acetylcholinesterase. Among these four laboratory strains and in individuals collected from natural populations, we found a correlation between increased acetylcholinesterase activities and higher dominance levels. We propose a molecular explanation for how variation in acetylcholinesterase activity may result in variation of dominance level. We also conjecture that the four resistant strains did not differ in their amino acid sequence in the catalytically active regions of acetylcholinesterase, but that the expression of the gene was regulated by either neighboring or distant sites, thereby modifying the dominance level. Under this interpretation, dominance levels may evolve in this system, since heritable variation in acetylcholinesterase activity was found.

Acetylcholinesterase↗

PubMed1996

Determination of Ace.1 Genotypes in Single Mosquitoes: Toward an Ecumenical Biochemical Test

The occurrence of two acetylcholinesterases, AChE1 and AChE2, in the mosquito Culex pipiens has been recently documented. Resistance to organophosphates and carbamates due to target insensitivity is the result of a qualitative change of only AChE1, encoded by the Ace.1 gene. Because AChE1 and AChE2 differ in their sensitivity to inhibitors, Ace.1 genotypes can be misclassified by previous tests. We describe a new rapid microplate test that allows unambiguous identification of Ace.1 genotypes. This test involves comparing AChE activities in the absence of insecticide and in the presence of two propoxur concentrations: a low concentration that inhibits only the sensitive AChE1 and a higher concentration that inhibits also AChE2 but not the insensitive AChE1 responsible of insecticide resistance. This comparison allows the identification of the three Ace.1 genotypes: resistant (Ace.1RR), susceptible (Ace.1SS) homozygotes, and heterozygotes (Ace.1RS). The similarity of propoxur sensitivity of modified AChE1s found in various resistant strains from the United States, Europe, and Africa indicates that this test is probably suitable for all the Ace.1 alleles described so far in C. pipiens.

Journal Article↗

PubMed1996

Duplication of the Ace.1 locus in Culex pipiens mosquitoes from the Caribbean.

In Culex pipiens mosquitoes, AChE1 encoded by the locus Ace.1 is the target of organophosphorus and carbamate insecticides. In several resistant strains homozygous for Ace.1RR, insensitive AChE1 is exclusively found. An unusual situation occurs in two Caribbean resistant strains where each mosquito, at each generation, displays a mixture of sensitive and insensitive AChE1. These mosquitoes are not heterozygotes, Ace.1RS, as preimaginal mortalities cannot account for the lethality of both homozygous classes. This situation is best explained by the existence of two Ace.1 loci, coding, respectively, a sensitive and an insensitive AChE1. Thus, we suggest that in the Caribbean a duplication of the Ace.1 locus occurred before the appearance of insecticide resistance at one of the two copies.

Acetylcholinesterase↗

PubMed1996

Existence of two acetylcholinesterases in the mosquito Culex pipiens (Diptera:Culicidae).

Two acetylcholinesterases (AChEs), AChE1 and AChE2, differing in substrate specificity and in some aspects of inhibitor sensitivity, have been characterized in the mosquito Culex pipiens. The results of ultracentrifugation in sucrose gradients and nondenaturing gel electrophoresis of AChE activity peak fractions show that each AChE is present as two molecular forms: one amphiphilic dimer possessing a glycolipid anchor and one hydrophilic dimer that does not interact with nondenaturing detergents. Treatment by phosphatidylinositol-specific phospholipase C converts each type of amphiphilic dimer into the corresponding hydrophilic dimer. Molecular forms of AChE1 have a lower electrophoretic mobility than those of AChE2. However, amphiphilic dimers and hydrophilic dimers have similar sedimentation coefficients (5.5S and 6.5S, respectively). AChE1 and AChE2 dimers, amphiphilic or hydrophilic, resist dithiothreitol reduction under conditions that allow reduction of Drosophila AChE dimers. In the insecticide-susceptible strain S-LAB, AChE1 is inhibited by 5 x 10(-4) M propoxur (a carbamate insecticide), whereas AChE2 is resistant. All animals are killed by this concentration of propoxur, indicating that only AChE1 fulfills the physiological function of neurotransmitter hydrolysis at synapses. In the insecticide-resistant strain, MSE, there is no mortality after exposure to 5 x 10(-4) M propoxur: AChE2 sensitivity to propoxur is unchanged, whereas AChE1 is now resistant to 5 x 10(-4) M propoxur. The possibility that AChE1 and AChE2 are products of tissue-specific posttranslational modifications of a single gene is discussed, but we suggest, based on recent results obtained at the molecular level in mosquitoes, that they are encoded by two different genes.

Acetylcholinesterase↗

PubMed1996

Dominance of insecticide resistance presents a plastic response.

Dominance level of insecticide resistance provided by one major gene (an insensitive acetylcholinesterase) in the mosquito Culex pipiens was studied in two distinct environments. Dominance level was found to be very different environments, varying from almost complete dominance to almost recessive when either propoxur (a carbamate insecticide) or chlorpyrifos (an organophosphorus insecticide) was used. To better understand this plastic response, three environmental parameters were manipulated and their interactions studied. For chlorpyrifos, each parameter had a small effect, but when all parameters were changed, the dominance level was greatly affected. For propoxur, one environmental parameter had a large effect by itself. It was further studied to understand the causal relationship of this plasticity. Recessivity of resistance was associated with more demanding environments. These results are discussed in the context of the various theories of the evolution of dominance. It appears that dominance of insecticide resistance cannot be directly predicted by Wright's physiological theory.

Animals↗