Avaliação biológica e biotecnológica de uma serpina (CfSerpina) do endoparasitoide Cotesia flavipes Cameron, 1891 (Hymenoptera: Braconidae) sobre o seu hospedeiro Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae)
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Data
2018-02-20
Autores
Silva, Juliana Barroso
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Universidade Estadual Paulista (Unesp)
Resumo
Os parasitoides são insetos que utilizam outros insetos para se desenvolverem e completarem seu ciclo imaturo de desenvolvimento. Os parasitoides podem manipular a fisiologia e afetar o crescimento e desenvolvimento de seus hospedeiros, o que frequentemente resulta na morte de suas vítimas. Durante o parasitismo de Cotesia flavipes Cameron, 1891 (Hymenoptera: Braconidae) sobre Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae), teratócitos são liberados após eclosão da larva do parasitoide no interior do hospedeiro e esses teratócitos produzem um transcrito que codifica uma serpina (CfSerpina). As serpinas são inibidores de serino proteases muitas vezes associadas à regulação da cascata de ativação de profenoloxidases em fenoloxidases, enzimas relacionadas com a melanização de corpos estranhos no hospedeiro. O presente estudo foi conduzido para investigar a função biológica da CfSerpina e avaliar seu potencial biotecnológico como agente de controle de D. saccharalis após ingestão. A CfSerpina foi produzida em sistema heterólogo em células de E. coli BL 21, purificada em coluna de níquel-sepharose e quantificada pelo método de Bradford para posterior detecção por Western blot. A proteína foi produzida em quantidades suficientes para a realização da avaliação biológica e biotecnológica. Após os ensaios, verificou-se que a CfSerpina não pode ser associada à inibição da ativação da cascata de profenoloxidases na hemolinfa de D. saccharalis. Além disso, a exploração biotecnológica da CfSerpina como uma proteína tóxica para D. saccharalis não causou mortalidade ou alterações no ganho de peso larval de D. saccharalis após ingestão e não resultou em inibição in vitro da atividade de proteases digestivas de D. saccharalis. Investigações sobre outras possibilidades de regulação da fisiologia de D. saccharalis por CfSerpina precisam ser conduzidos para definição do papel dessa proteína no processo de regulação hospedeira.
Parasitoids are insects which use other insects to develop and complete their immature cycle. Parasitoids can manipulate the physiology and affect the growth and development of their hosts, which often results in the death of their victims. During parasitism of Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) by the parasitoid Cotesia flavipes Cameron, 1891 (Hymenoptera: Braconidae) teratocytes are released after the eclosion of parasitoid larvae inside the host and these teratocytes produce a transcript that encodes a putative serpin (CfSerpin). Serpins are serine proteases inhibitors often associated with the regulation of the activation of prophenoloxidases into phenoloxidases, enzymes responsible for melanization of foreign bodies in the host. The study was conducted to investigate the biological function of CfSerpin, as well to evaluate its biotechnological potential to disrupt the development of D. saccharalis larvae after ingestion. CfSerpin was produced in a heterologous system using E. coli BL 21 cells, purified on a nickel-sepharose column, quantified by the Bradford method, and detected by Western blot. Purified CfSerpin was produced in sufficient amount for biological and biotechnological evaluation. CfSerpin could not be associated with the inhibition of activation of prophenoloxidases cascade in the host insect hemolymph. In addition, biotechnological exploitation of CfSerpin as a toxic protein after ingestion did not resulted in the disruption of the development of D. saccharalis larvae and no in vitro inhibition of digestive proteases of D. saccharalis by CfSerpin was observed. Investigations to unravel the role of CfSerpin on the regulation of D. saccharalis physiology still need to be conducted.
Parasitoids are insects which use other insects to develop and complete their immature cycle. Parasitoids can manipulate the physiology and affect the growth and development of their hosts, which often results in the death of their victims. During parasitism of Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) by the parasitoid Cotesia flavipes Cameron, 1891 (Hymenoptera: Braconidae) teratocytes are released after the eclosion of parasitoid larvae inside the host and these teratocytes produce a transcript that encodes a putative serpin (CfSerpin). Serpins are serine proteases inhibitors often associated with the regulation of the activation of prophenoloxidases into phenoloxidases, enzymes responsible for melanization of foreign bodies in the host. The study was conducted to investigate the biological function of CfSerpin, as well to evaluate its biotechnological potential to disrupt the development of D. saccharalis larvae after ingestion. CfSerpin was produced in a heterologous system using E. coli BL 21 cells, purified on a nickel-sepharose column, quantified by the Bradford method, and detected by Western blot. Purified CfSerpin was produced in sufficient amount for biological and biotechnological evaluation. CfSerpin could not be associated with the inhibition of activation of prophenoloxidases cascade in the host insect hemolymph. In addition, biotechnological exploitation of CfSerpin as a toxic protein after ingestion did not resulted in the disruption of the development of D. saccharalis larvae and no in vitro inhibition of digestive proteases of D. saccharalis by CfSerpin was observed. Investigations to unravel the role of CfSerpin on the regulation of D. saccharalis physiology still need to be conducted.
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Regulação hospedeira, teratócitos, cenobionte, função biológica, biotecnologia, Host regulation, teratocytes, koinobiont, biological function, biotechnology