Download Functional Characterization of Soybean Transcription Factor

Functional Characterization of Soybean Transcription Factor GmbZIPE2
Gonçalves, A.B.; Alves, M.S.; Dadalto, S.P.; Fietto L.G.
Dep. de Bioquímica e Biologia Molecular, UFV, Viçosa, MG, Brazil.
INTRODUCTION: Transcription factors are proteins that regulate gene expression
by binding to specific sequences in DNA. Transcription factors are among the
major targets to increase the tolerance of plants to stresses, since these proteins
control the expression of several genes simultaneously. Members of the bZIP
family of transcription factors are characterized by having a leucine zipper domain
which is a basic DNA binding domain. Previous research demonstrated that the
GmbZIPE2 gene in soybean (Glyma05g30170.1) is responsive to pathogens .
OBJECTIVES: The objective of this study was functionally characterize the
transcription factor GmbZIPE2, by discovering which promoters binds to it, as well
as determining their possible modulation of target genes. MATERIAL AND
METHODS: In this research, GmbZIPE2 was amplified and cloned into a bacterial
expression vector (pDEST17) and this was inserted into Escherichia coli strain
C41 in which the recombinant protein was successfully expressed. The
GmbZIPE2 protein was used in electrophoretic mobility shift assay (EMSA) using
two cis-elements related to defense against pathogens, H-box and G-box.
qRT-PCR experiments were performed to establish the relationship between the
overexpression of GmbZIPE2 and expression of genes related to plant defense
against pathogens. RESULTS: Our results demonstrated that GmbZIPE2 binds to
H-box, but does not bind to the G-box. It was found that overexpression of
GmbZIPE2 in soybean protoplasts does not interfere with the expression of
GmPAL-1 and GmOMT (genes encoding proteins related to the biosynthesis of
lignin and phenylpropanoid), but it activates the expression of
pathogenesis-related gene 1 (GmPR1) gene which expression is induced in
response to a variety of pathogens. CONCLUSIONS: These results suggest that
GmbZIPE2 modulates the expression of genes related to defense against
pathogens in plants.
Word Keys: biotic stress, bZIP, Glycine max
Supported by: CAPES, CNPq and FAPEMIG