Multiple resistance to pirimiphos-methyl and bifenthrin in Tribolium castaneum involves the activity of lipases, esterases, and laccase2.
Several recent studies have elucidated the molecular mechanisms that confer insecticide resistance on insect pests. However, little is known about multiple resistance in red flour beetle (Tribolium castaneum) at molecular level. The multiple resistance is characterized as resistance to different classes of insecticides that have different target sites, and is mediated by several enzymatic systems. In this study, we investigated the biochemical and molecular mechanisms involved in multiple resistance of T. castaneum to bifenthrin (pyrethroid [Pyr]) and pirimiphos-methyl (organophosphate [Org]). We used artificial selection, biochemical and in silico approaches including structural computational biology. After five generations of artificial selection in the presence of bifenthrin (F5Pyr) or pirimiphos-methyl (F5Org), we found high levels of multiple resistance. The hierarchical enzymatic cluster revealed a pool of esterases (E), lipases (LIPs) and laccase2 (LAC2) potentially contributing to the resistance in different ways throughout development, after one or more generations in the presence of insecticides. The enzyme-insecticide interaction network indicated that E2, E3, LIP3, and LAC2 are enzymes potentially required for multiple resistance phenotype. Kinetic analysis of esterases from F5Pyr and F5Org showed that pirimiphos-methyl and specially bifenthrin promote enzyme inhibition, indicating that esterases mediate resistance by sequestering bifenthrin and pirimiphos-methyl. Our computational data were in accordance with kinetic results, indicating that bifenthrin has higher affinity at the active site of esterase than pirimiphos-methyl. We also report the capability of these insecticides to modify the development in T. castaneum. Our study provide insights into the biochemical mechanisms employed by T. castaneum to acquire multiple resistance.
Departamento de Biotecnologia, Genética e Biologia Celular da Universidade Estadual de Maringá, Paraná, Brazil. Electronic address: alison_sheep@hotmail.com.
Alison Henrique Ferreira
AH
Julio
Departamento de Biotecnologia, Genética e Biologia Celular da Universidade Estadual de Maringá, Paraná, Brazil.
Adriana Aparecida Sinópolis
AA
Gigliolli
Centro de Ciências Exatas Departamento de Ciências, Universidade Estadual de Maringá Campus de Umuarama, Paraná, Brazil.
Kátia Aparecida Kern
KA
Cardoso
Departamento de Biotecnologia, Genética e Biologia Celular da Universidade Estadual de Maringá, Paraná, Brazil.
Sandro Daniel
SD
Drosdoski
Departamento de Biotecnologia, Genética e Biologia Celular da Universidade Estadual de Maringá, Paraná, Brazil.
Rodrigo Amaral
RA
Kulza
Departamento de Bioquímica da Universidade Estadual de Maringá, Paraná, Brazil.
Flávio Augusto Vicente
FA
Seixas
Departamento de Biotecnologia, Genética e Biologia Celular da Universidade Estadual de Maringá, Paraná, Brazil.
Maria Claudia Colla
MC
Ruvolo-Takasusuki
Departamento de Bioquímica da Universidade Estadual de Maringá, Paraná, Brazil.
Cristina Giatti Marques
CG
de Souza
Departamento de Biotecnologia, Genética e Biologia Celular da Universidade Estadual de Maringá, Paraná, Brazil. Electronic address: aslapenta@uem.br.
Ana Silvia
AS
Lapenta
D000818:Animals; D018432:Drug Resistance, Multiple / Q000187:drug effects*; D004591:Electrophoresis, Polyacrylamide Gel; D004950:Esterases / Q000145:classification / Q000235:genetics / Q000378:metabolism; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007305:Insecticide Resistance / Q000187:drug effects*; D007306:Insecticides / Q000633:toxicity; D007527:Isoenzymes / Q000145:classification / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D042845:Laccase / Q000145:classification / Q000235:genetics / Q000378:metabolism; D008049:Lipase / Q000145:classification / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D009946:Organothiophosphorus Compounds / Q000633:toxicity*; D010802:Phylogeny; D000072417:Protein Domains; D011679:Pupa / Q000033:anatomy & histology / Q000187:drug effects / Q000201:enzymology; D011722:Pyrethrins / Q000633:toxicity*; D014231:Tribolium / Q000033:anatomy & histology / Q000201:enzymology* / Q000378:metabolism
D019476:Insect Proteins; D007306:Insecticides; D007527:Isoenzymes; D009946:Organothiophosphorus Compounds; D011722:Pyrethrins; C014153:pirimiphos methyl; C099952:bifenthrin; D042845:Laccase; D004950:Esterases; D008049:Lipase
Artificial selection; Enzymatic inhibition; Interaction network; Juvenile hormone esterase; Organophosphate; Pesticide resistance; Pyrethroid; Red flour beetle