The tomato borer Tuta absoluta (Lepidoptera: Gelechiidae) is an invasive pest of tomato crops that is rapidly expanding around the world. It is considered a devastating pest and its control heavily relies on application of insecticides. Diamides are a novel class of insecticides acting on insect ryanodine receptors and are highly effective against lepidopteran pests. To date, chlorantraniliprole and flubendiamide have been registered in the market and they have been extensively used to manage T. absoluta. In this study, a survey was conducted in Greece and Italy monitoring diamide resistance. The populations originating from Sicily (Italy) exhibited LC 50 s that ranged between 47.6-435 for chlorantraniliprole and 993-1.376 for flubendiamide, while for Crete (Greece) LC 50 s ranged between 0.14-2.45 for chlorantraniliprole and 1.7-8.4 for flubendiamide (LC 50 s in mg L -1 ). Comparing this result to the susceptible reference strain, high resistance levels for the Italian populations were detected, i.e., up to 2,414-and 1,742-fold for chlorantraniliprole and flubendiamide, respectively. Resistance ratios for Greek populations were found up to 14-fold for chlorantraniliprole and 11-fold for flubendiamide, suggesting that diamide resistance is low but increasing considering monitoring data over time. Hereby, we report for the first time, cases of resistance development to diamide insecticides in T. absoluta. These findings underline the importance of committing to the resistance management strategies for diamide insecticides.
Bemisia tabaci resistance evolved differently in each of the three insecticides studied. Imidacloprid resistance seems less established and less persistent than alpha-cypermethrin resistance. The low resistance levels for pirimiphos-methyl suggest absence of cross-resistance with other organophosphates or carbamates used.
We investigated pyrethroid resistance mechanisms in Tetranychus urticae strains from Greece. Combined bioassay, biochemical and synergistic data indicated that although P450 mono-oxygenase activities were associated with the trait, target site insensitivity was the major resistance component. A 3.3 kb cDNA fragment of the T. urticae para sodium channel gene encompassing segment 4 of domain II to segment 6 of domain IV was obtained by a degenerate PCR strategy. The T. urticae sequence showed highest identity (56%) to the scabies mite, Sarcoptes scabiei, and was phylogenetically classified within the divergent group of Arachnida. Comparison of resistant and susceptible strains identified the point mutation F1538I in segment 6 of domain III, which is known to confer strong resistance to pyrethroids, along with a second mutation (A1215D) in the intracellular linker connecting domains II and III with an unknown role. Three transcripts were identified corresponding to the k and l alternative exons. The mode of inheritance of resistance was confirmed as incompletely recessive, which is consistent with a target site mechanism for pyrethroids.
Four AChE1 mutations were found in resistant strains of T. urticae, and three of them, F331W, G328A and A201S, are possibly involved in resistance to OP and CARB insecticides. Among them, F331W is probably the most important and the most common in T. urticae. It can be easily detected by the diagnostic PCR-RLFP assay developed in this study.
We investigated the molecular diversity of the major agricultural pest Bemisia tabaci and of its associated secondary endosymbionts in Greece. Analyzing mitochondrial DNA, we found that the Q1 (=Q west) is predominant. We used eight microsatellite polymorphic markers to study the genetic structure of 37 populations from mainland and insular Greece, collected on different host species from outdoor and protected crops as well as from non-cultivated plants. In some cases, gene flow was found to be low even between populations separated by just a few kilometres. Bayesian analysis identified two main genetic groups, the first encompassing populations from south Crete and the second composed of populations from north Crete, two other Aegean islands and mainland Greece. Genetic differentiation was not correlated with different host plant species or habitat, or greenhouse versus open environment populations. Gene flow significantly decreased with geographic distance, but no isolation by distance existed when only the samples from mainland Greece or only the samples from Crete were considered. The secondary symbionts Wolbachia and Hamiltonella were present at high frequencies while Arsenophonus, Cardinium and Rickettsia were absent from Greek populations. Multilocus sequence typing of Wolbachia identified two Wolbachia strains. These two strains were found together in most of the populations studied but never in the same host individual. Their role on the observed population structure is discussed.
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