Isolation and Expression Analysis of DFR-A Gene from Petunia hybtida

doi:10.11924/j.issn.1000-6850.2013-2373

Abstract

Abstract: After isolation the DFR-A from Petunia hybrida, its expression characteristic and the DFR enzyme activity in different tissues were studied. It will be helpful for the isolation and study of other anthocyanin structural genes. Based on the petunia DFR gene sequences published by NCBI, the full length cDNA of DFR was isolated from the petals of petunia. Real-time quantitative PCR (qRT-PCR) was performed to determine the expression pattern of DFR in different tissues of petunia. The DFR enzyme activities in different tissues were measured also. (1) 1473 bp cDNA sequence of petunia DFR- A was cloned. Bioinformatics analysis showed that the open reading frame of this gene was 1143 bp, encoding 380 amino acids. (2) Expression analysis showed that the expression of DFR gene was detected in all tissues of Petunia. But the expression levels were different; the expression level of this gene was highest in anther, medium in petal of flower starting to open and flower fully opened, lower in flower buds, and the lowest expression level of DFR was in leaf. (3) DFR activity analysis revealed that DFR was almost no activity in leaf and anther, low activity in flower buds, the highest activity in flower starting to open. (4) There was correlation between DFR activity and DFR mRNA concentration except in anther. The DFR enzyme activity of Petunia was mainly controlled by the transcriptional regulation.

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