Abstract

Bruchid resistance is regulated by defensin gene. Vigna radiata plant defensin 1 (VrPDF1) inhibits alpha-amylase activities in insect gut; therefore, insect will die of undigestion starch. VrPDF1 content in mungbean seeds is very low. Hence an increase in content of VrPDF1 in mungbean seeds leads to enhance alpha -amylase inhibition in larvae and bruchids, which is necessary for the study to improve bruchid resistance in mungbeans. This article presents the results of VrPDF1 gene expression in T1 generation transgenic tobacco seeds. It was confirmed that VrPDF1 gene was attached to genome of tobacco plants and translated to synthesize VrPDF1. Recombinant VrPDF1 protein was successfully expressed in seeds of four transgenic tobacco lines (T1-7, T1-8, T1-10, T1-11), among which T1-10 line had the highest recombinant VrPDF1 content, reaching 8.57 g mg-1 total protein. The extract containing recombinant VrPDF1 from the transgenic tobacco lines effectively inhibited the activity of alpha-amylase from the intestine of larvae and weevils. However, the protein extract solution from T1-10 line had the strongest inhibitory effect, so the activity of alpha-amylase was only 18.89% compared to controls. The analysis results of VrPDF1 gene expression in transgenic tobacco plants are fundamental to the transfer of pPhaso-dest-VrPDF1 vector into mungbean to improve bruchid resistance of mungbean and contribute to improving mungbean preservation.

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