A novel PR10 promoter from Erianthus arundinaceus directs high constitutive transgene expression and is enhanced upon wounding in heterologous plant systems

• DOI: 10.1007/s11033-015-3934-2
• Publication Date: 2015-12-15T05:57:19Z
• Authors: M. Chakravarthi; Divya P. Syamaladevi; P. Harunipriya; Sruthy Maria Augu...; N. Subramonian
• Keywords: Nicotiana; 0301 basic medicine; 2. Zero hunger; Base Sequence; Oryza; Cyclopentanes; Acetates; Regulatory Sequences, Nucleic Acid; Plants, Genetically Modified; Saccharum; Plant Leaves; 03 medical and health sciences; Gene Expression Regulation, Plant; Genes, Reporter; Oxylipins; Transgenes; Promoter Regions, Genetic; Abscisic Acid; Plant Proteins

In genetic engineering, inducible promoters play an important role as the expression of genes driven by them can be turned on or off under situations like biotic or abiotic factors. There are few reports on inducible promoters that can be employed in the development of transgenic plants, particularly in sugarcane. In the present study, four wound inducible genes (Chitinase, PR1A, PR10 and HRGP) were selected and were amplified from Erianthus arundinaceus, a distant relative of sugarcane. In order to determine the gene that is highly induced upon wounding, RT-qPCR was performed, which showed that PR10 gene expression was instantaneous and higher upon wounding when compared to the other three genes. Using the random amplification of genomic ends technique, a 592 bp promoter sequence was obtained and in silico analysis of the upstream regulatory region revealed a 469 bp promoter and 123 bp of 5' untranslated region (UTR). Functional analyses of the promoter sequence (with and without 5' UTR) in tobacco, rice and sugarcane using β-glucuronidase (GUS) as the reporter gene revealed the constitutive and inducible nature of the PR10 promoter. Our studies have demonstrated that the PR10 promoter, though highly constitutive, was quickly induced upon wounding as well as on treatment with abscisic acid and methyl jasmonate hormones. This is the first report on the isolation and characterization of a PR10 promoter from a wild grass and is expected to have application for development of transgenic plants.