Keywords
Sequencing
Lepidoptera
Cry1Ac toxin
Bioinsecticide
How to Cite
Abstract
Anticarsia gemmatalis Hübner, 1818 is the main soybean defoliating pest in Brazil. The biological control of the species is done with products based on toxins produced by Bacillus thurigiensis (Bt), as bioinsecticides, or in transgenic plants. After activation by intestinal proteases, these toxins interact with receptors, especially cadherin, leading to death due to the formation of cellular pores. In recent years resistant populations have been identified in the laboratory, which can be a problem if the same patterns are found in crops, reducing their control effect. In this paper, we performed a comparative structural analysis of a mutation region for the gene of this receptor in A. gemmatalis, among resistant and susceptible strains treated with a toxin produced by Bt (Cry1Ac). The HaCad fragment of the cadherin gene was amplified by PCR, sequenced, and analyzed by bioinformatics tools. The PCR results were positive for resistant specimens but not for susceptible strains, suggesting the presence of a mutation in the resistant strain. In the sequenced fragments of the resistant insects, six haplotypes were found, and the originated amino acid sequences demonstrated the modification in four sites, which did not interfere with the three-dimensional shape of the protein. These data showed considerable variation taking into account the size of the fragment, even if they do not affect the final structure of the protein. The results allowed a better understanding of the mechanisms of resistance to Cry1Ac in the species, mainly in the involvement of cadherin in this process.
References
CONAB - Companhia Nacional de Abastecimento (Orgs). Acompanhamento da safra brasileira grãos. Observação Agrícola 2019; 6(9): 1-117.
Vieira ECS, Ávila CJ, Vivan LM, da Silva IF, Vieira MCS, Silva PG. Control of Anticarsia gemmatalis (Hübner: 1818) (Lepidoptera: Erebidae) and Chrysodeixis includens (Walker: 1858) (Lepidoptera: Noctuidae) through insecticides applied to soybean seeds. J Agric Sci 2019; 11(18): 88-97. https://doi.org/10.5539/jas.v11n18p88
Praça LB, Batista AC, Martins ÉS, Siqueira CB, Dias DGDS, Gomes ACMM, et al. Estirpes de Bacillus thuringiensis efetivas contra insetos das ordens Lepidoptera, Coleoptera e Diptera. Pesq Agropec Bras 2004; 39(1): 11-16. https://doi.org/10.1590/S0100-204X2004000100002.
Campagne P, Smouse PE, Pasquet R, Silvain JF, Le Ru B, Van den Berg J. Impact of violated high‐dose refuge assumptions on evolution of Bt resistance. Evol Appl 2016; 9(4): 596-607. https://doi.org/10.1111/eva.12355
Tabashnik BE, Carrière Y. Surge in insect resistance to transgenic crops and prospects for sustainability. Nat Biotechnol 2017; 35(10): 926. https://doi.org/10.1038/nbt.3974.
Kukanur VS, Singh TVK, Kranthi K, Andow D. Cry1Ac resistance allele frequency in field populations of Helicoverpa armigera (Hübner) collected in Telangana and Andhra Pradesh, India. Crop Prot 2018; 107: 34-40. https://doi.org/10.1016/j.cropro.2018.01.008
Gómez I, Miranda-Rios J, Rudiño-Piñera E, Oltean DI, Gill SS, Bravo A, Soberón M. Hydropathic complementarity determines interaction of epitope 869HITDTNNK876 in Manduca sexta Bt-R1 receptor with loop 2 of domain II of Bacillus thuringiensis Cry1A toxins. J Biol Chem 2002; 277(33): 30137-30143. https://doi.org/10.1074/jbc.M203121200.
Zhang H, Wu S, Yang Y, Tabashnik BE, Wu Y. Non-recessive Bt toxin resistance conferred by an intracellular cadherin mutation in field- selected populations of cotton bollworm. PLOS One 2012; 7(12): 53418. https://doi.org/10.1371/journal.pone.0053418
Greene GL, Leppla NC, Dickerson WA. Velvetbean Caterpillar: A rearing procedure and artificial medium. J Econ Entomol 1976; 69(4): 487-488. https://doi.org/10.1093/jee/69.4.487
Suzuki KM, Arias MC, Giangarelli DC, Freiria GA, Sofia SH. Mitochondrial DNA diversity of orchid bee Euglossa fimbriata (Hymenoptera: Apidae) populations assessed by PCRRFLP. Biochem Genet 2010; 48(3-4): 326-341. http://doi.org/ 10.1007/s10528-009-9325-4
Xiao Y, Dai Q, Hu R, Pacheco S, Yang Y, Liang G, et al. A single point mutation resulting in cadherin mislocalization underpins resistance against Bacillus thuringiensis toxin in cotton bollworm. J Biol Chem 2017; 292(7): 2933-2943. https://doi.org/10.1074/jbc.M116.768671
Costa-da-Silva A, Capurro M, Bracco J. Genetic lineages in the yellow fever mosquito Aedes (Stegomyia) aegypti (Diptera: Culicidae) from Peru. Mem Inst Oswaldo Cruz 2005; 100(6): 539-544. https://doi.org/10.1590/S0074-02762005000600007
Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 2016; 33(7): 1870-1874. https://doi.org/10.1093/molbev/msw054.
Hall T. Bioedit v 7.0.5 Biological sequences alignment editor for Windows. Ibis Therapeutics, a division of Isis Pharmaceuticals: Carlsbad 2005.
Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 1994; 22(22): 4673-4680. https://doi.org/10.1093/nar/22.22.4673
Excoffier L, Lischer HE. Arlequin suite ver 3.5: a new series of programs to perform population genetics analyses under Linux and Windows. Mol Ecol Resour 2010; 10(3): 564-567. https://doi.org/10.1111/j.1755-0998.2010.02847.x.
Waterhouse A, Bertoni M, Bienert S, Studer G, Tauriello G, Gumienny R, Lepore R. SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res 2018; 46(1): 296-303. https://doi.or/10.1093/nar/gky427
Melo AL, Soccol VT, Soccol CR. Bacillus thuringiensis: mechanism of action, resistance, and new applications: a review. Crit Rev Biotechnol 2016; 36(2): 317-326. https://doi.org/10.3109/07388551.2014.960793.
Wang L, Wang J, Ma Y, Wan P, Liu K, Cong S, Li X. Transposon insertion causes cadherin mis-splicing and confers resistance to Bt cotton in pink bollworm from China. Sci Rep 2019; 9(1): 7479. https://doi.org/10.1038/s41598-019-43889-x
Wang L, Ma Y, Want P, Liu K, Xiao Y, Wang J, Li X. Resistance to Bacillus thuringiensis linked with a cadherin transmembrane mutation affecting cellular trafficking in pink bollworm from China. Insect Biochem Mol Biol 2018; 94: 328-354. https://doi.org/10.1016/j.ibmb.2018.01.004
Pezenti LF, Sosa-Gómez DR, De Souza RF, Vilas-Boas LA, Gonçalves KB, Da Silva CRM, et al. Transcriptional profiling analysis of susceptible and resistant strains of Anticarsia gemmatalis and their response to Bacillus thuringiensis. Genomics 2021; 113: 2264-2275.
Alberts B, Johnson A, Raff M, Lewis J, Roberts K, Walter P, et al. Biologia molecular da célula, sixth ed. Artmed: Porto Alegre 2017.
Bel Y, Escriche B. Common genomic structure for the Lepidoptera cadherin-like genes. Gene 2006; 381: 71-80. https://doi.org/10.1016/j.gene.2006.07.001
McGaughey W, Gould F, Gelernter W. Bt resistance management. Nat Biotechnol 1998; 16(2): 144-146. https://doi.org/10.1038/nbt0298-144.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Copyright (c) 2022 Felipe Cordeiro Dias, Jaqueline Fernanda Dionísio, Daniel Ricardo Sosa-Gómez, Renata da Rosa