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

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• 作者：Railene de Azevedo Pereira ; Joã ; o Aguiar Nogueira Batista ; Maria Cristina Mattar da Silva ; Osmundo Brilhante de Oliveira Neto ; Edson Luiz Zangrando Figueira ; Arnubio Valencia Jimé ; nez and Mari
• 关键词：Coffee berry borer ; Phaseolus coccineus ; Digestive enzymes ; Insect
• 刊名：Phytochemistry
• 出版年：2006
• 出版时间：September 2006
• 年：2006
• 卷：67
• 期：18
• 页码：2009-2016
• 全文大小：433 K

文摘

Plant α-amylase inhibitors are proteins found in several plants, and play a key role in natural defenses. In this study, a gene encoding an α-amylase inhibitor, named αAI-Pc1, was isolated from cotyledons of Phaseolus coccineus. This inhibitor has an enhanced primary structure to P. vulgaris α-amylase inhibitors (α-AI1 and α-AI2). The αAI-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 αAI-1, while no reaction was observed with untransformed tobacco plants. Immunological assays showed that the αAI-Pc1 gene product represented up to 0.05 % of total soluble proteins in T0 plants seeds. Furthermore, recombinant αAI-Pc1 expressed in tobacco plants was able to inhibit 65 % of digestive H. hampei α-amylases. The data herein suggest that the protein encoded by the αAI-Pc1 gene has potential to be introduced into coffee plants in order to increase their resistance to the coffee berry borer.