title: Proteomic profile of hemolymph and detection of induced antimicrobial peptides in response to microbial challenge in Diatraea saccharalis (Lepidoptera: Crambidae).
abstract: Insects are organisms extremely well adapted to diverse habitats, primarily due to their innate immune system, which provides them with a range of cellular and humoral responses against microorganisms. Lepidoptera hemolymph proteins involved in humoral responses are well known; however, there is a lack of knowledge about the sugarcane borer Diatraea saccharalis. In this present work, the hemolymph proteins of this pest insect were studied by applying proteomic methodologies. Two-dimensional electrophoresis (2-DE) gels of proteins extracted from naive larvae and larvae challenged with Escherichia coli (ATCC 11224) and Bacillus subtilis (ATCC 6623) showed an average of 300 spots, and 92 of these spots corresponded in all three 2-DE gels. Forty-one spots were excised and digested with trypsin and analyzed using mass spectrometry. After analysis, 10 proteins were identified, including some proteins of the immune system: β-defensin-like protein, Turandot A-like protein, attacin-like protein, peptidoglycan recognition protein and cyclophilin-like protein. Nine proteins were present in both experimental conditions; however, β-defensin-like protein was present only in hemolymph challenged by B. subtilis. Notably, attacin-like protein was strongly induced by challenge with E. coli, suggesting an immune response against the infection. However, antimicrobial activity was observed in the test zone of microbial growth inhibition of B. subtilis solely with the hemolymph extract of the larvae challenged with B. subtilis. We made for the first time a proteomic profile of the hemolymph of D. saccharalis in which it was possible to identify the presence of important proteins involved in the immune response.
authors.affiliation: Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, Cascavel, Paraná, Brazil.
authors.forename: Iara Fernanda
authors.initials: IF
authors.lastname: Rocha
authors.affiliation: Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, Cascavel, Paraná, Brazil.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Maller
authors.affiliation: Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, Cascavel, Paraná, Brazil.
authors.forename: Rita
authors.initials: R
authors.lastname: de Cássia Garcia Simão
authors.affiliation: Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, Cascavel, Paraná, Brazil.
authors.forename: Marina Kimiko
authors.initials: MK
authors.lastname: Kadowaki
authors.affiliation: Centro de Ciências Biológicas e da Saúde, Universidade Estadual do Oeste do Paraná, Cascavel, Paraná, Brazil.
authors.forename: Luis Francisco
authors.initials: LF
authors.lastname: Angeli Alves
authors.affiliation: Departamento de Bioquímica, Universidade Federal do Paraná, Curitiba, Paraná, Brazil.
authors.forename: Luciano Fernandes
authors.initials: LF
authors.lastname: Huergo
authors.affiliation: Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, Cascavel, Paraná, Brazil. Electronic address: jlcsilva@gmail.com.
authors.forename: José Luis
authors.initials: JL
authors.lastname: da Conceição Silva
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000032:analysis* / Q000378:metabolism; D001412:Bacillus subtilis / Q000502:physiology; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism; D004926:Escherichia coli / Q000502:physiology; D006458:Hemolymph / Q000382:microbiology* / Q000502:physiology; D054884:Host-Pathogen Interactions; D019476:Insect Proteins / Q000032:analysis* / Q000378:metabolism; D007915:Lepidoptera / Q000382:microbiology* / Q000502:physiology; D040901:Proteomics
chemical_list: D023181:Antimicrobial Cationic Peptides; D002352:Carrier Proteins; D019476:Insect Proteins; C118897:peptidoglycan recognition protein
keywords: Antimicrobial peptide; Diatraea saccharalis; Hemolymph; Proteome