No abstract

Biological control requires specific tools for the accurate detection and identification of natural enemies, and to detect unusual variations in their density, which may follow changes in agricultural practices. Here we have developed specific molecular markers to detect Lydella thompsoni (Herting) and Pseudoperichaeta nigrolineata (Walker) (Diptera: Tachinidae) within the European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae). Primers amplifying fragments of the mitochondrial COI gene were designed following alignment of comparable sequences for a range of parasitoid and host species. Each of the primer pairs proved to be species specific to a tachinid species, amplifying DNA fragments of 191 and 91 bp in length for L. thompsoni and P. nigrolineata, respectively. This DNA-based technique allowed molecular evaluation of parasitism in O. nubilalis natural populations. In order to study the geographical distribution of both species in France, O. nubilalis diapausing larvae in maize stalks were collected from 12 locations over the whole country. The molecular evaluation of parasitism was compared with the traditional method of maintaining O. nubilalis populations in controlled conditions before breaking off the diapause. The percentage parasitism found in both species of tachinids was higher--approximately three times--using the molecular method, suggesting an underestimation by the traditional rearing protocol. Tachinid parasitism on O. nubilalis was not significantly different between geographical areas (south, central and north France) for both species. This study shows that molecular methods are very promising for the correct detection and identification of tachinid parasitoids in natural field populations.

-Mycotoxins produced by Fusarium spp. during plant cultivation induce severe diseases in animal and humans. In 2007 a European Union regulation set maximum concentrations of mycotoxins in maize and derivatives of 4000 ppb for fumonisins B 1 and B 2 , 1750 ppb for deoxynivalenol, and 350 ppb for zearalenone. To assess the safety of French maize food, investigations are currently being carried out by the national Biological Risk Monitoring network. Here, 84 plots were cropped with the Bt maize MON 810 and its isogenic non-Bt counterpart in 2005 and 2006 in Southwestern France. Mycotoxin levels were measured in grain at harvest. Fumonisins B 1 and B 2 , deoxynivalenol, and zearalenone were analysed by liquid chromatography-mass spectrometry. The data were analysed statistically using non-parametric tests for mycotoxins and analysis of variance for weather variables. As the climate was homogenous inside the experimental area, the transgenic event introduced into the maize was the only key parameter which differed between Bt and non-Bt maize plots. Our results show that Bt maize decreased concentrations of fumonisins by 90% and zearalenone by 50%, whereas the concentration of deoxynivalenol was slightly increased. Those findings suggest a competition among Fusarium species that produce fumonisins or trichothecenes. According to the European regulation, 93% of the Bt maize crops can be sold, compared with only 45% for non-Bt maize plots. Our results thus show that Bt maize improved food safety by greatly reducing mycotoxin levels in field crops in Southwestern France.Bt MON810 maize / fumonisins B 1 and B 2 / deoxynivalenol / zearalenone / Regulation 1126/2007

Considering the growing trade of seed potato, reliable diagnostic protocols are required for the detection of regulated nematode species. In this study, a specific and sensitive multiplex Taqman-based real-time PCR method was developed in order to detect and identify Globodera pallida, G. rostochiensis and Heterodera schachtii. The newly designed primers and probes enabled the detection of all the target populations tested and with no cross-reaction for closely related non-target species (55 populations tested). The limit of detection (LOD) was one juvenile for G. rostochiensis and G. pallida and five juveniles for H. schachtii. For monitoring potato cyst nematodes, this analytical tool would extend the number of cyst investigated as five juveniles can be detected among 50 cysts in a sample. Furthermore, this multiplex assay detects DNA of the three targeted species in template DNA obtained directly from float material after nematode extraction from soil.