Abstract
Bacillus thuringiensis (Bt) is a biological alternative to the indiscriminate use of chemical insecticides in agriculture. Due to resistance development on insect pests to Bt crops, isolating novel Bt strains is a strategy for screening new pesticidal proteins or strains containing toxin profile variety that can delay resistance. Besides, the combined genomic and proteomic approaches allow identifying pesticidal proteins and virulence factors accurately. Here, the genome of a novel Bt strain (Bt TOL651) was sequenced, and the proteins from the spore–crystal mixture were identified by proteomic analysis. Toxicity bioassays with the spore–crystal mixture against larvae of Diatraea saccharalis and Anticarsia gemmatalis, key pests of sugarcane and soybean, respectively, were performed. The toxicity of Bt TOL651 varies with the insect; A. gemmatalis (LC50 = 1.45 ng cm−2) is more susceptible than D. saccharalis (LC50 = 73.77 ng cm−2). Phylogenetic analysis of the gyrB gene indicates that TOL651 is related to Bt kenyae strains. The genomic analysis revealed the presence of cry1Aa18, cry1Ac5, cry1Ia44, and cry2Aa9 pesticidal genes. Virulence factor genes such as phospholipases (plcA, piplc), metalloproteases (inhA), hemolysins (cytK, hlyIII, hblA, hblC, hblD), and enterotoxins (nheA, nheB, nheC) were also identified. The combined use of the genomic and proteomic data indicated the expression of Cry1Aa18, Cry1Ac5, and Cry2Aa9 proteins, with Cry1Ac5 being the most abundant. InhA1 also was expressed and may contribute to Bt TOL651 pathogenicity. These results provide Bt TOL651 as a new tool for the biocontrol of lepidopteran pests.
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This work was supported by grants from the National Council of Scientific and Technological Development (CNPq—process numbers: 313455/2019-8; 427304/2018-0; 163178/2020-8), Tocantins State Foundation for Research Aid (FAPT-SESAU/TO-DECIT/SCTIE/MS_CNPQ/N° 01/2017) and Federal University of Tocantins (PROPESQ)—EDITAL Nº 29/2020 PROPESQ and PPGBIOTEC/UFT/GURUPI- Chamada pública para auxílio de tradução e/ou publicação de artigos científicos-EDITAL Nº 011/2020.
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RWSA, GBA, and EEO designed the experiments. GBA, RWSA, and BMR performed the genomic and proteomic data analysis. GBA, LOVJ, GRS, BMR, and RWSA drafted the manuscript. RWSA, EEO, LOVJ, BMR, MAO, and MMS revised the manuscript. All authors read and approved the final manuscript.
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Alves, G.B., de Oliveira, E.E., Jumbo, L.O.V. et al. Genomic–proteomic analysis of a novel Bacillus thuringiensis strain: toxicity against two lepidopteran pests, abundance of Cry1Ac5 toxin, and presence of InhA1 virulence factor. Arch Microbiol 205, 143 (2023). https://doi.org/10.1007/s00203-023-03479-y
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DOI: https://doi.org/10.1007/s00203-023-03479-y