Relationship between toxicity of Vip3Aa proteins and their binding capacity to intestine receptors of lepidopteran pestsAbstract -The objective of this work was to evaluate the toxicity of new Vip3Aa proteins and their binding capacity to brush-border membrane vesicles (BBMV) in the intestine of Spodoptera frugiperda, Anticarsia gemmatalis, and Heliothis virescens neonate larvae. The proteins expressed by the genes vip3Aa42 and vip3Aa43 showed toxicity to S. frugiperda (LC 50 of 78.2 and 113 ng cm -2, respectively) and A. gemmatalis (LC 50 of 239.2 and 57.5 ng cm -2 , respectively), but they showed low toxicity to H. virescens (LC 50 >5,000 ng cm -2 ). BBMV binding assays showed that the proteins bind effectively to the receptors on vesicles of the evaluated species, but this binding capacity is only effective on the activation of toxicity to the evaluated populations of S. frugiperda and A. gemmatalis.
A bactéria Bacillus thuringiensis Berliner produz um corpo de inclusão paraesporal (cristal) de natureza proteica, formado durante a esporulação, que atua de forma eficiente no controle de insetos-praga de culturas economicamente importantes. Esse cristal é constituído de proteínas Cry, que são codificadas pelos genes cry; um isolado pode ser caracterizado pelo conteúdo de genes cry que apresenta. Visando caracterizar novos isolados no combate de insetos-praga pertencentes às ordens Lepidoptera e Coleoptera, 76 isolados bacterianos foram analisados molecularmente e tiveram seu potencial de controle avaliado por meio de bioensaios com larvas de Spodoptera frugiperda (J.E. Smith), Sphenophorus levis Vaurie e Tenebrio molitor Linnaeus. As análises moleculares indicaram 11 isolados (14,5% da coleção), contendo genes lepidóptero-específicos e 17 (22,37%) com genes coleóptero-específicos. As análises de patogenicidade revelaram dois isolados com alto potencial de controle para lagartas de S. frugiperda, um para larvas de S. levis e seis prejudiciais ao desenvolvimento das larvas de T. molitor. Esses isolados de B. thuringiensis podem ser promissores no controle biológico das referidas pragas.
Palavras-chave: controle biológico, genes cry, Spodoptera frugiperda, Sphenophorus levis, Tenebrio molitor.
Characterization of new strains of Bacillus thuringiensis for the control of important insect pests in agriculture
AbstractBacillus thuringiensis Berliner produces protein crystals that act effectively on the control of pests for several relevant crops. The cry genes codes for these proteins are named Cry proteins. The cry gene profile can potentially set each bacterial isolate as a potential pest control agent. Aiming to analyze new isolates for the controlling of pests belonging to the orders Lepidoptera and Coleoptera, 76 samples were molecularly analyzed and had their potential for control evaluated through bioassays with larvae of Spodoptera frugiperda (J.E. Smith), Sphenophorus levis Vaurie and Tenebrio molitor Linnaeus. Based on molecular analysis data, 11 samples (14.5%) were able to control lepidopteran specific pests due to their cry genes content, and 17 samples (22.37%) exhibited coleopteran specific cry genes. The bioassays results indicated two samples with highly effective against S. frugiperda, one against S. levis and six with high toxicity against T. molitor larvae development. The selected B. thuringiensis isolates may be promising for the biological control of these pests.
O objetivo deste trabalho foi identificar e caracterizar os genes cry3, vip1, vip2 e vip1/vip2 em uma coleção de 1.078 isolados de Bacillus thuringiensis potencialmente tóxicos para larvas de coleópteros. Foram utilizados pares de oligonucleotídeos iniciadores gerais obtidos a partir de regiões conservadas dos genes e do alinhamento de sequências consenso. Posteriormente, os isolados positivos foram caracterizados por meio da técnica de PCR‑RFLP, tendo-se utilizado enzimas de restrição específicas, para identificar novas subclasses de genes nos isolados. Cento e cinquenta e um isolados foram positivos para os genes avaliados, com maior frequência para o gene vip1/vip2 (139 isolados). Pela técnica de PCR‑RFLP, foram observados 14 perfis polimórficos, o que indica a presença de diferentes alelos e, consequentemente, de distintas subclasses desses genes.
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