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Entada acaciifolia trypsin inhibitor: structural characterization and evaluation of insecticide potential against the fall armyworm (Spodoptera frugiperda)

Grant number: 11/09361-0
Support Opportunities:Regular Research Grants
Duration: October 01, 2011 - September 30, 2013
Field of knowledge:Biological Sciences - Biology
Principal Investigator:Sergio Marangoni
Grantee:Sergio Marangoni
Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Associated researchers: Caio Fernando Ramalho de Oliveira


The search for alternative insecticide compounds that don't present the risks associated to the chemical insecticides now used is object of research of several groups and an alternative for a maintainable agriculture. Among the different groups of plant proteins with insecticide activity known, we can detach the class of the proteinases inhibitors as one of the more studied. The objectives of this project involve the accomplishment of a meticulous study of the insecticide potential of EATI, a new trypsin inhibitor isolated from Entada acaciifolia seeds, on Spodoptera frugiperda, pest insect responsible for severe damages in different crops in the national scenery. One second segment of this project seeks to acquire knowledge concerning the structure of this inhibitor, through studies of sequential homology, three-dimensional modelling for homology and SAXS studies, correlating their structural characteristics the function that carries out. The inhibitor EATI will already be purified starting from seeds of E. acaciifolia through a methodology established, based on different chromatographic techniques. Since then, its insecticide potential will be analyzed on the larval development, pupal and of emergency of the adult insects. For so much, different concentrations of the inhibitor will be introduced in artificial diet and analyzed at the end of each stage. Besides the direct effect of EATI on the reduction of the weight and survival in all of the stages of the development cycle, several nutritional parameters will be obtained as well as the accomplishment of rehearsals in vitro with extracted digestive enzymes of the gut of the insects. The differential genic expression of the larval enzymes fed in diet controls and in the presence of the inhibitor EATI will be analyzed through RT-PCR, in order to determine modifications in the expression of enzymes induced by the exhibition to the inhibitor in study, determining its insecticide mechanism. The structural focus of the project seeks the obtaining of a three-dimensional model built through studies of sequential homology, accomplished in platforms of HPLC associated to the MALDI-TOF MS and SAXS.Based on the results of this project hoped to contribute with information that can be used as model and to suggest the job of this inhibitor as a biotechnological tool in the improvement of plants attacked by this pest. (AU)

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Scientific publications (4)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
RAMALHO DE OLIVEIRA, CAIO FERNANDO; MARANGONI, SERGIO; RODRIGUES MACEDO, MARIA LIGIA. The trypsin inhibitor from Entada acaciifolia seeds affects negatively the development of Mediterranean flour moth, Anagasta kuehniella. PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY, v. 108, p. 74-79, . (11/09361-0)
RAMALHO DE OLIVEIRA, CAIO FERNANDO; SOUZA, THAIS DE PAULA; POSTALI PARRA, JOSE ROBERTO; MARANGONI, SERGIO; SILVA-FILHO, MARCIO C.; RODRIGUES MACEDO, MARIA LIGIA. Insensitive trypsins are differentially transcribed during Spodoptera frugiperda adaptation against plant protease inhibitors. COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, v. 165, n. 1, p. 19-25, . (11/09361-0)
DANTZGER, MIRIAM; VASCONCELOS, ILKA MARIA; SCORSATO, VALERIA; APARICIO, RICARDO; MARANGONI, SERGIO; RODRIGUES MACEDO, MARIA LIGIA. Bowman-Birk proteinase inhibitor from Clitoria fairchildiarta seeds: Isolation, biochemical properties and insecticidal potential. Phytochemistry, v. 118, p. 224-235, . (11/09361-0)
RAMALHO DE OLIVEIRA, CAIO FERNANDO; VASCONCELOS, ILKA MARIA; APARICIO, RICARDO; MACHADO FREIRE, MARIA DAS GRACAS; BALDASSO, PAULO APARECIDO; MARANGONI, SERGIO; RODRIGUES MACEDO, MARIA LIGIA. Purification and biochemical properties of a Kunitz-type trypsin inhibitor from Entada acaciifolia (Benth.) seeds. Process Biochemistry, v. 47, n. 6, p. 929-935, . (11/09361-0)

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