Genetic diversity of Brazilian Bacillus thuringiensis isolates with toxicity against Aedes aegypti (Diptera: Culicidae).

Bacillus thuringiensis (Bt) isolates native to Maranhão (BtMA) that are highly toxic to Aedes aegypti larvae and seven standard subspecies of Bt were analyzed for genetic diversity using the rep-PRC technique with BOX, ERIC, REP, MB1, and GTG5 markers. The rep-PCR technique is considered an extremely reliable, reproducible, fast and highly discriminatory technique that may be used even among populations of the same species. These five markers revealed a total of 38 polymorphic DNA fragments for 30 BtMA isolates. Eight groups were obtained with the dendrogram generated through Pearson's correlation analysis, with four groups formed only with BtMA isolates and four comprised of isolates of BtMA and the standard subspecies toxic to dipterans and lepidopterans. Despite the high genetic diversity of BtMA, a low correlation between the collection site, gene content and mortality against A. aegypti larvae was evidenced. The clustering of the standard subspecies of Bt that were toxic against dipterans with BtMA isolates confirm the mosquitocidal action of the native isolates from Maranhão, and they can be used as an alternative for A. aegypti control and other insects of medical importance and for the control of agricultural pests.

Inheritance and Fitness of Plutella xylostella (Lepidoptera: Plutellidae) Resistance to Chlorfenapyr.

The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is a key pest of Brassicaceae worldwide. Populations have globally evolved resistance to various insecticides including chlorfenapyr, which was observed at high frequency in Brazil. We report the genetic characterization and fitness costs associated with chlorfenapyr resistance in a field-derived strain. The resistant strain (BZR-RR) and a susceptible strain (REC-SS) were used in both concentration-response bioassays and demography-based approach. Inheritance pattern of chlorfenapyr resistance was determined by conducting reciprocal crosses between susceptible and resistant strains, and by backcrossing. Next, life table analysis for the susceptible, heterozygotes, and resistant strains was performed to assess eventual fitness costs associated with chlorfenapyr resistance. Resistance of P. xylostella (BZR-RR) strain to chlorfenapyr was very high (RR50 = 421.58-fold) and also autosomal (no differences between reciprocal crosses), monofactorial and incompletely dominant (F1 pool DD = 0.26 ± 0.14). Dominance (h) was concentration dependent with 16 mg/l allowing at least 95% survival of the resistant heterozygotes. Recessive fitness cost was observed to be associated with resistance to chlorfenapyr. The relative fitness of heterozygotes (RS) and resistant homozygotes (BZR-RR) in comparison to the susceptible strain (REC-SS) was 0.91 and 0.23, respectively. Significant differences were found for many fitness components in the resistant homozygotes. Altogether, results suggest a rational use of chlorfenapyr in areas where susceptible populations still prevail, in parallel with the use of diagnostic concentrations (e. g., 20 mg chlorfenapyr/l), and rotation with different mode of actions, for which fitness costs of resistance are nonrecessive in P. xylostella.

Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane.

The biological potential of Vip and Cry proteins from Bacillus is well known and widely established. Thus, it is important to look for new genes showing different modes of action, selecting those with differentiated entomotoxic activity against Diatraea flavipennella and Elasmopalpus lignosellus, which are secondary pests of sugarcane. Therefore, Cry1 and Vip3 proteins were expressed in Escherichia coli, and their toxicities were evaluated based on bioassays using neonate larvae. Of those, the most toxic were Cry1Ac and Vip3Aa considering the LC50 values. Toxins from E. coli were purified, solubilized, trypsinized, and biotinylated. Brush Border Membrane Vesicles (BBMVs) were prepared from intestines of the two species to perform homologous and heterologous competition assays. The binding assays demonstrated interactions between Cry1Aa, Cry1Ac, and Vip3Aa toxins and proteins from the BBMV of D. flavipennella and E. lignosellus. Homologous competition assays demonstrated that binding to one of the BBMV proteins was specific for each toxin. Heterologous competition assays indicated that Vip3Aa was unable to compete for Cry1Ac toxin binding. Our results suggest that Cry1Ac and Vip3Aa may have potential in future production of transgenic sugarcane for control of D. flavipennella and E. lignosellus, but more research is needed on the potential antagonism or synergism of the toxins in these pests.

Insecticidal activity against Bemisia tabaci biotype B of peel essential oil of Citrus sinensis var. pear and Citrus aurantium cultivated in northeast Brazil.

The fumigant action of peel essential oils of Citrus sinensis var. pear (pear orange = PO) and C. aurantium (bitter orange = BO) from the northeast of Brazil were evaluated against Bemisia tabaci biotype B and compared with eugenol as a positive control. The oil concentration in the PO at 8.5 microL/L of air caused 97% mortality, while the oil concentration of BO at 9.5 microL/L of air caused 99% mortality. However, the LC50 estimates for both oils (LC50 = 3.80 microL/L of air for PO and LC50 = 5.80 microL/L of air for BO) did not differ from each other, but they did when compared with eugenol (LC50 = 0.20 microL/L of air). Regarding their effects on oviposition, the Citrus oils showed concentration-response dependence, reducing the number of eggs as the concentration increased, which was not observed for eugenol. The minimum concentrations of the oils that caused a significant reduction in the egg lay were 3.5 and 7.0 microL/L of air for BO and PO, respectively. These results suggest that oils from PO and BO peels may be promising as models to develop new insecticides that might be applied into the integrated management of whiteflies.