RESUMO
AIMS: To demonstrate the ability of Bacillus thuringiensis to penetrate as spore-crystal complex to the internal tissues of bean plants, and keep its insecticidal activity. To test the vertical transmission of the spore-crystal complex in Arabidopsis thaliana. METHODS AND RESULTS: The experimental strain was transformed with the pMUTIN-gfp plasmid which labelled the spores of B. thuringiensis HD-73 with the GFP protein. Once the rhizosphere of the bean plants was inoculated with the labelled strain, the bacterium was recovered from leaves, stems, and petioles. Furthermore, toxicity of treated plants was significantly higher than control plants when bio-assayed on cabbage looper larvae. The labelled strain was recovered from the dead insects. When the rhizosphere of A. thaliana plants was inoculated with the labelled strain, mature seeds from these plants were surface-sterilized and grown under in vitro conditions. The labelled strain was recovered from the seedlings. CONCLUSIONS: We showed that B. thuringiensis subsp. kurstaki (HD-73) in the rhizosphere can translocate to upper tissues of bean plants, and keep its insecticidal activity. Transmission of the labelled B. thuringiensis strain passed to the next generation of A. thaliana. SIGNIFICANCE AND IMPACT OF THE STUDY: The role of B. thuringiensis as a potential facultative endophyte bacterium and the possible biotechnological repercussions are discussed.
Assuntos
Arabidopsis/microbiologia , Bacillus thuringiensis/fisiologia , Proteínas de Bactérias/toxicidade , Endotoxinas/toxicidade , Proteínas Hemolisinas/toxicidade , Inseticidas , Phaseolus/microbiologia , Animais , Bacillus thuringiensis/genética , Bacillus thuringiensis/isolamento & purificação , Toxinas de Bacillus thuringiensis , Proteínas de Bactérias/análise , Proteínas de Bactérias/genética , Endotoxinas/análise , Endotoxinas/genética , Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/genética , Proteínas Hemolisinas/análise , Proteínas Hemolisinas/genética , Inseticidas/análise , Larva/microbiologia , Mariposas/crescimento & desenvolvimento , Mariposas/microbiologia , RizosferaRESUMO
Previous observations on high virulence of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Trichoplusia ni single nucleopolyhedrovirus (TnSNPV) acting together led us to test possible synergism between these two nucleopolyhedroviruses (NPVs) on cabbage looper larvae. Because synergism between AcMNPV and the Trichoplusia ni granulovirus (TnGV) has been well established before, these two viruses were included in this study as a positive control. Each virus was assayed separately on first-instar cabbage looper and their LC50s were estimated at 2.33, 0.39 and 462 OB/mm2 diet for AcMNPV, TnSNPV and TnGV, respectively. LC50s of AcMNPV mixed with sub-lethal concentrations of TnSNPV and TnGV increased 8 and 10.7 times, respectively. Synergism between the viruses was analyzed by the ANOVA test for the LC50s, the Plackett and Hewlett's joint-action rate test, and the Tammes-Bakuniak graphic method. All three analyses corroborated the synergism between the viruses. The presence of a putative enhancin in the TnSNPV was analyzed by Southern blot hybridization, using a 1.5 kbp KpnI fragment from the TnGV vef gene as a probe. No hybridization was observed. The occurrence of a new putative synergistic factor in TnSNPV is discussed.