Faten Raboudi

Use of molecular and In Silico bioinformatic tools to investigate pesticide binding to insect (Lepidoptera) phenoloxidases (PO): Insights to toxicological aspects

In the present work, the promising bioinformatic tools, based on structure–affinity analysis, allowed to screen several pesticides supposed to bind to the insect immune Phenoloxidases (PO). First, the binding of aminoparathion, a reference compound, to the PO was structurally (3D) validated in accordance with previous reports. Second, using the same docking conditions, a range of pesticides was checked for their binding ability to the crystal 3D structure (PDB 3HSS) of the insect Manduca sexta (Lepidoptera) PO. The obtained data showed that many of the tested pesticides were able to bind, in silico, to M. sexta PO. The combination of in vitro (chemical and biochemical) and in silico (automated docking) approaches was found advantageous to elucidate the behavior of phenolic pesticides as substrate-analogues when binding to the active site of insect POs. Our findings emphasize new ecotoxicological aspects of pesticide residues in the agro-chemical and environmental circles.

A novel method for molecular targeting of insecticide resistance in Rhopalosiphum padi L. (Homoptera: Aphididae)

ABSTRACT The bird cherry-oat aphid (Rhopalosiphum padi L.) is a devastating cereal pest that develops high resistance to organophosphate and carbamate insecticides. Because acetylcholinesterase (ACE) is the target of carbamate and organophosphate insecticides, the resistance mechanism usually involves mutations occurring in ACE-encoding genes, Ace1 and Ace2. Here, we describe a novel polymerase chain reaction (PCR)-based method for the diagnosis of resistance cases associated with the point mutation F368L in the Ace2 gene. We amplified a 127 bp DNA fragment from Ace2 gene using a modified reverse primer, and digested the amplification product using SmaI endonuclease. This procedure enabled a simple and rapid distinction between resistant and susceptible genotypes for F368L mutation. Subsequently, we screened 152 R. padi samples, and found that F368L mutation occurred at low frequency, in both the homozygous (R/R) and heterozygous (R/S) states. Based upon the results of this study, we believe that molecular diagnosis of insecticide resistance should be generalized to genes and mutations involved in this process, toward an optimal accuracy of insecticide applications.

Spatial and Temporal Genetic Variation in Tunisian Field Populations of Macrosiphum euphorbiae (Thomas)

ABSTRACT The potato aphid Macrosiphum euphorbiae (Thomas) is a major pest of several economic crops in Tunisia. Using 10 microsatellites, we analyzed five populations of M. euphorbiae sampled during the 2004–2005 solanaceous and cucurbit season (April through September) from five geographic origins. From 235 aphids, 61 different multilocus genotypes were identified of which three genotypes MLG1, MLG2, and MLG42 were predominant on all host plants and regions. MLG1 and MLG2 genotypes were detected in 2004 and did not reappear in spring 2005, while the genotype MLG42 was detected only in 2005. All populations showed significant deviation from Hardy—Weinberg equilibrium even in data sets including one individual per genotype, &khgr; 2 independence tests and analysis of molecular variance showed no significant differentiation among populations collected on different host plant and from different geographic origin, but differences between populations from two successive years were significant. Factorial component analysis corroborates these results. The probable causes of this seasonal variation were discussed.

Structural Modeling of Plant Secondary MetabolitesCharacterized by an Inhibitory Potential of Insect Enzymes

Structural Modeling of Plant Secondary Metabolites Characterized by an Inhibitory Potential of Insect Enzymes Management of pests in the world is facing economic and environmental challenges posed by the majority of chemical pesticides. The identification of new effective insecticides is essential to fight against these parasites. Inhibition of insect enzymes has been opined as a possible mode of action of various plant secondary metabolites. However, it is necessary to gain knowledge about the binding mode of these metabolites to the target region so as to facilitate an understanding of the evolution of novel molecules for pest management.