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A polymorphic (GA/CT)n- SSR influences promoter activity of Tryptophan decarboxylase gene in Catharanthus roseus L. Don

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Title A polymorphic (GA/CT)n- SSR influences promoter activity of Tryptophan decarboxylase gene in Catharanthus roseus L. Don
 
Creator Kumar, Santosh
Bhatia, Sabhyata
 
Subject Plant biotechnology
Plant molecular biology
Transcriptional regulatory elements
 
Description Accepted date: 17 August 2016
Simple Sequence Repeats (SSRs) of polypurine-polypyrimidine type motifs occur very frequently in the 5′ flanks of genes in plants and have recently been implicated to have a role in regulation of gene expression. In this study, 2 accessions of Catharanthus roseus having (CT)8 and (CT)21 varying motifs in the 5′UTR of Tryptophan decarboxylase (Tdc) gene, were investigated for its role in regulation of gene expression. Extensive Tdc gene expression analysis in the 2 accessions was carried out both at the level of transcription and translation. Transcript abundance was estimated using Northern analysis and qRT-PCR, whereas the rate of Tdc gene transcription was assessed using in-situ nuclear run-on transcription assay. Translation status of Tdc gene was monitored by quantification of polysome associated Tdc mRNA using qRT-PCR. These observations were validated through transient expression analysis using the fusion construct [CaM35S:(CT)8–21:GUS]. Our study demonstrated that not only does the length of (CT)n -SSRs influences the promoter activity, but the presence of SSRs per se in the 5′-UTR significantly enhances the level of gene expression. We termed this phenomenon as “microsatellite mediated enhancement” (MME) of gene expression. Results presented here will provide leads for engineering plants with enhanced amounts of medicinally important alkaloids.
We acknowledge the National Institute of Plant Genome Research (NIPGR), New Delhi, India for the funding support and the Council of Scientific and Industrial Research (CSIR) for fellowship grant to SK.
 
Date 2016-09-16T11:34:57Z
2016-09-16T11:34:57Z
2016
 
Type Article
 
Identifier Scientific Reports, 6: 33280
2045-2322
http://59.163.192.83:8080/jspui/handle/123456789/681
http://www.nature.com/articles/srep33280
10.1038/srep33280
 
Language en_US
 
Publisher Nature Publishing Group