Transient Assay System for the Analysis ofPR-1aGene Promoter in Tobacco BY-2 Cells

Abstract

In order to develop a rapid and versatile assay system suitable for the analysis of regulated expression of tobacco pathogenesis-related protein 1a (PR-1a) gene, we investigated the use of the transient gene expression system in tobacco BY-2 cells by microprojectile bombardment. Using dual luciferase assay as a reporter gene expression detection system, we observed significant induction of PR-1a promoter activity by salicylic acid (SA) treatment. On the other hand, treatment with 4-hydroxybenzoic acid (4-HBA) resulted in no detectable increase in luciferase activity. Co-expression of a trans-acting factor, the NPR1/NIM1 protein of Arabidopsis, resulted in the induction of higher expression levels of the PR-1a promoter. These results suggest that the assay system is applicable for the analysis of factors involved in the regulated expression of SA-inducible defense-related genes.

Regulation of Copper Homeostasis by Micro-RNA in Arabidopsis

“…When restriction sites are not indicated in primers, the site present in the genome sequence was used for the cloning. The amplified product was digested with XbaI and NcoI, then cloned into the pBI221-LUCϩ vector (29), and confirmed by sequencing. The primers used for PCR amplification were CSD2ox-F (5Ј-TTCCTCTA-GACGTCAAACATAGC-3Ј) and CSD2cut3-R (5Ј-CTAACA-ACATGACCCATGGAGCTCCAGAA-3Ј) for CSD2⌬1::LUC, CSD2ox-F and CSD2cut2-R (5Ј-CATAAATGCCATGGC-CGATGGT-3Ј) for CSD2⌬2::LUC, and CSD2ox-F and CSD2full-R2 (5Ј-CTTTCGCAGGTGTGATTGCCATGGCG-3Ј) for CSD2full::LUC.…”

A model system to screen for candidate plant activators using an immune-induction system in Arabidopsis

“…The tobacco PR-1a gene, which is strictly upregulated after activation of the SA-dependent signal transduction pathway, has been used as a marker to monitor chemical activation of this pathway, and transgenic tobacco BY-2 cells harboring the PR-1a promoter-luciferase fusion gene have been suggested to be useful for the study of defense gene expression (Ono et al 2004). This system may also be used to screen for plant activators after modification for high through-put screening procedures (Ono et al 2004;Ogura et al 2005).…”

GmWRKY53, a water- and salt-inducible soybean gene for rapid dissection of regulatory elements in BY-2 cell culture

“…luciferase as a reporter, 5 for example stably transformed BY-2 cells have successfully been used to dissect JA-inducible promoters. 6,7 In addition, this BY-2 system can also be extended to the testing of individual promoter elements and synthetic promoters that respond to JA.…”