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Effect of soybean peptidase inhibitors in the pattern of gene expression and gut enzyme activity of larvae of Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) Given the importance of search for more sustainable approaches for pest control, plant peptidases inhibitors emerge as a promising tool for the protection of crops by obtaining transgenic plants. Harmful effects of soybean peptidases inhibitors (SPI) intake by Diatraea saccharalis on its development were already reported in previous studies, however, these effects have not been studied at the molecular level for this specie. This study aimed to identify and analyze the gene expression and the gut serine peptidases activity of D. saccharalis in response to the intake of an active extract of soybean peptidases inhibitors. These informations were related to the presence or absence of some adaptive mechanism of this insect in response to these inhibitors. The gut transcriptome of larvae of D. saccharalis in the fifth developmental instar was obtained, undergoing chronic intake of IPS. Then, sequences of serino peptidases like trypsin and chymotrypsin were identified and used for gene expression studies on a real-time PCR in response to acute (for 24 and 48 hours) and chronic intake of 0.5 % (w/v) of SPI active extract by larvae of D. saccharalis in the third and fifth instars of larval development. The same sequences were used to propose phylogenies, including comparisons with sequences of Spodoptera frugiperda serine peptidases identified in another parallel study. Tryptic and chymotryptic activity assays were performed using specific substrates (BApNA and SApNA) to determine the effect of the SPI intake on enzyme activity and finally, the integrity and the inhibitory activity of the SPI were analyzed after passing through the digestive tract of caterpillars. The results showed that under the experimental conditions employed in this study, the larvae of D. saccharalis were not able to modulate the gene expression of serine peptidases in response to acute or chronic SPI intake. The results of tryptic and chymotryptic activity due to the SPI intake can suggest a synergism in the action of these enzymes in larvae of the fifth developmental instar. SPI intake did not cause differences in enzymatic activity of third instar larvae. Electrophoretic separation of proteins from the feces of caterpillars enabled visualization of a band with a molecular mass around 20 kDa possibly corresponding to the Kunitz inhibitor present in soybean. The antitryptic activity of the feces indicates that D. saccharalis is not able to hydrolyze soybean peptidases inhibitor as a mechanism of adaptation to their presence in the diet.

Efeito dos inibidores de peptidase de soja no padrão de expressão e atividade enzimática intestinal de lagartas de Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae)