An a-amylase inhibitor gene from Phaseolus coccineus encodes a protein with potential for control of coffee berry borer (Hypothenemus hampei)

• Autor(a) principal: Pereira, Railene de Azevedo
• Data de Publicação: 2006
• Outros Autores: Batista, João Aguiar Nogueira, Silva, Maria Cristina Mattar da, Oliveira Neto, Osmundo Brilhante de, Figueira, Edson Luiz Zangrando, Jiménez, Arnubio Valencia, Grossi-de-Sá, Maria Fátima
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UCB
• Texto Completo: http://twingo.ucb.br:8080/jspui/handle/10869/675; https://repositorio.ucb.br:9443/jspui/handle/123456789/7810
• topic: Coffee berry borer; Phaseolus coccineus; Digestive enzymes; Insect

Plant a-amylase inhibitors are proteins found in several plants, and play a key role in natural defenses. In this study, a gene encoding an a-amylase inhibitor, named aAI-Pc1, was isolated from cotyledons of Phaseolus coccineus. This inhibitor has an enhanced primary structure to P. vulgaris a-amylase inhibitors (a-AI1 and a-AI2). The aAI-Pc1 gene, constructed with the PHA-L phytohemaglutinin promoter, was introduced into tobacco plants, with its expression in regenerated (T0) and progeny (T1) transformant plants monitored by PCR amplification, enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis, respectively. Seed protein extracts from selected transformants reacted positively with a polyclonal antibody raised against aAI-1, while no reaction was observed with untransformed tobacco plants. Immunological assays showed that the aAI-Pc1 gene product represented up to 0.05% of total soluble proteins in T0 plants seeds. Furthermore, recombinant aAI-Pc1 expressed in tobacco plants was able to inhibit 65% of digestive H. hampei a-amylases. The data herein suggest that the protein encoded by the aAI-Pc1 gene has potential to be introduced into coffee plants in order to increase their resistance to the coffee berry borer.