Long-term studies on the evolution of resistance of Myzus persicae (Hemiptera: Aphididae) to insecticides in Greece.

The aphid Myzus persicae s.l. (Hemiptera: Aphididae) is an important pest of many crops worldwide with a complex life cycle, intensely controlled by chemical pesticides, and has developed resistance to almost all used insecticides. In Greece, the aphid exhibits high genetic variation and adaptability and it is a classic example of evolution in the making. We have been studying M. persicae for over 20 years, on different host plants and varying geographical areas, analyzing its bio-ecology and the ability to develop resistance to insecticides. In this review, we present new and historical data on the effectiveness of insecticides from seven chemical groups used to control the aphid in Greece and the incidence of seven resistance mechanisms, including the new fast-spreading R81T point mutation of the postsynaptic nicotinic acetylcholine receptor. Thousands of samples were tested by biological, biochemical and molecular assays. The aphid populations were found to have developed and maintain resistance at medium to high levels to organophosphates, carbamates, pyrethroids and neonicotinoids for decades. In the latter group, a marked increase is recorded during an ~10-year period. The data analyzed and the extensive bibliography, advocate the difficulty to control the aphid making the design and application of IPM/IRM programs a challenge. We discuss principles and recommendations for the management of resistance, including the use of compounds such as flonicamid, spirotetramat, flupyradifurone and sulfoxaflor. We emphasize that resistance is a dynamic phenomenon, changing in time and space, requiring, therefore, continuous monitoring.

A small deletion in the olive fly acetylcholinesterase gene associated with high levels of organophosphate resistance.

Organophosphate resistance in the olive fly was previously shown to associate with two point mutations in the ace gene. The frequency of these mutations was monitored in Bactrocera oleae individuals of increasing resistance. In spite of the difference in resistance among the individuals, there was no correlation between mutation frequencies and resistance level, indicating that other factors may contribute to this variation. The search for additional mutations in the ace gene of highly resistant insects revealed a small deletion at the carboxyl terminal of the protein (termed Delta3Q). Significant correlation was shown between the mutation frequency and resistance level in natural populations. In addition, remaining activity of acetylcholinesterase enzyme (AChE) after dimethoate inhibition was higher in genotypes carrying the mutation. These results strongly suggest a role of Delta3Q in high levels of organophosphate (OP) resistance. Interestingly, the carboxyl terminal of AChE is normally cleaved and substituted by a glycosylphosphatidylinositol (GPI) anchor. We hypothesize that Delta3Q may improve GPI anchoring, thus increasing the amount of AChE that reaches the synaptic cleft. In this way, despite the presence of insecticide, enough enzyme would remain in the cleft for its normal role of acetylcholine hydrolysis, allowing the insect to survive. This provides a previously un-described mechanism of resistance.

Genetic variation among Mediterranean populations of Sesamia nonagrioides (Lepidoptera: Noctuidae) as revealed by RFLP mtDNA analysis.

Restriction fragment length polymorphism analysis of two segments of mitochondrial DNA (COI and 16S rRNA) was used to examine genetic variation in Sesamia nonagrioides (Lefèbvre) populations from the Mediterranean basin. Four populations were collected from central and southern Greece, and five from northern latitudes: Greece, Italy, France and Spain. No variation was observed in COI, while 16S rRNA segment proved highly polymorphic and 28 different haplotypes were found. Lower intra-population polymorphism was found in the northern populations than in southern ones. Although no significant isolation by distance was found, the UPGMA tree based on Nei's raw number of nucleotide differences separated the populations into two major groups, i.e. one with the northern (40.6 degrees N-43.4 degrees N) and the other with the southern populations (37.3 degrees N-39.2 degrees N). Analysis of molecular variance revealed that most of the variation was between the two major groups (Phi(CT)=0.559), and all pairwise comparisons between the northern and southern populations resulted in high and significant F(ST) values (overall F(ST)=0.604). The high F(ST) values and the strong spatial genetic structure indicate that long-distance migration may be a rare event. The populations do not seem to have experienced a strong historical bottleneck. The occurrence of a few widespread haplotypes and the genetic similarity of the northern populations could be attributed to a historical expansion of certain haplotypes from the south towards to the northern borders of the species' distribution area.