title: Toxicity to cotton boll weevil Anthonomus grandis of a trypsin inhibitor from chickpea seeds.
abstract: Cotton (Gossypium hirsutum L.) is an important agricultural commodity, which is attacked by several pests such as the cotton boll weevil Anthonomus grandis. Adult A. grandis feed on fruits and leaf petioles, reducing drastically the crop production. The predominance of boll weevil digestive serine proteinases has motivated inhibitor screenings in order to discover new ones with the capability to reduce the digestion process. The present study describes a novel proteinase inhibitor from chickpea seeds (Cicer arietinum L.) and its effects against A. grandis. This inhibitor, named CaTI, was purified by using affinity Red-Sepharose Cl-6B chromatography, followed by reversed-phase HPLC (Vydac C18-TP). SDS-PAGE and MALDI-TOF analyses, showed a unique monomeric protein with a mass of 12,877 Da. Purified CaTI showed significant inhibitory activity against larval cotton boll weevil serine proteinases (78%) and against bovine pancreatic trypsin (73%), when analyzed by fluorimetric assays. Although the molecular mass of CaTI corresponded to alpha-amylase/trypsin bifunctional inhibitors masses, no inhibitory activity against insect and mammalian alpha-amylases was observed. In order to observe CaTI in vivo effects, an inhibitor rich fraction was added to an artificial diet at different concentrations. At 1.5% (w/w), CaTI caused severe development delay, several deformities and a mortality rate of approximately 45%. These results suggested that CaTI could be useful in the production of transgenic cotton plants with enhanced resistance toward cotton boll weevil.
authors.affiliation: Universidade Católica de Brasília, Pós-Graduação em Ciências Genômicas e Biotecnologia, SGAN Quadra 916, Módulo B, Av. W5 Norte 70.790-160-Asa Norte Brasília/DF, Brazil.
authors.forename: Angélica
authors.initials: A
authors.lastname: de P G Gomes
authors.forename: Simoni C
authors.initials: SC
authors.lastname: Dias
authors.forename: Carlos
authors.initials: C
authors.lastname: Bloch
authors.forename: Francislete R
authors.initials: FR
authors.lastname: Melo
authors.forename: José R
authors.initials: JR
authors.lastname: Furtado
authors.forename: Rose G
authors.initials: RG
authors.lastname: Monnerat
authors.forename: Maria F
authors.initials: MF
authors.lastname: Grossi-de-Sá
authors.forename: Octávio L
authors.initials: OL
authors.lastname: Franco
mesh_terms: D000383:Agriculture / Q000379:methods; D000818:Animals; D029863:Cicer / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D007814:Larva / Q000187:drug effects / Q000201:enzymology / Q000502:physiology; D008970:Molecular Weight; D010575:Pesticides / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000633:toxicity*; D012639:Seeds / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014361:Trypsin Inhibitors; D048889:Weevils / Q000187:drug effects* / Q000201:enzymology* / Q000254:growth & development / Q000502:physiology; D000516:alpha-Amylases / Q000037:antagonists & inhibitors
chemical_list: D010575:Pesticides; D010940:Plant Proteins; D014361:Trypsin Inhibitors; D000516:alpha-Amylases